U.S. patent number 8,592,576 [Application Number 13/123,327] was granted by the patent office on 2013-11-26 for unsymmetrical pyrrolobenzodiazepine-dimers for treatment of proliferative diseases.
This patent grant is currently assigned to Spirogen Sarl. The grantee listed for this patent is Stephen John Gregson, Philip Wilson Howard, Luke Masterson. Invention is credited to Stephen John Gregson, Philip Wilson Howard, Luke Masterson.
United States Patent |
8,592,576 |
Howard , et al. |
November 26, 2013 |
**Please see images for:
( Certificate of Correction ) ** |
Unsymmetrical pyrrolobenzodiazepine-dimers for treatment of
proliferative diseases
Abstract
Claimed are unsymmetrical Pyrrolobenzodiazepine-dimers
(PBD-dimers) of formula (I), that are unsymmetrical due at least to
the values that the variables R.sup.12 and R.sup.2 represent.
Formula (I), wherein: R.sup.2 is of formula (II), where A is a
C.sub.5-7 aryl group, X is selected from the group comprising: OH,
SH, CO.sub.2H, COH, N.dbd.C.dbd.O, NHR.sup.N, wherein R.sup.N is
selected from the group comprising H and C.sub.1-4 alkyl, and
(OC.sub.2H.sub.4).sub.mOCH.sub.3, where m is 1 to 3, and either:
(i) Q.sup.1 is a single bond, and Q.sup.2 is selected from a single
bond and --Z--(CH.sub.2).sub.n--, where Z is selected from a single
bond, O, S and NH and n is from 1 to 3; or (ii) Q.sup.1 is
--CH.dbd.CH--, and Q.sup.2 is a single bond; R.sup.12 is a
C.sub.5-10 aryl group, optionally substituted by one or more
substituents selected from the group comprising: halo, nitro,
cyano, ether, C.sub.1-7 alkyl, C.sub.3-7 heterocyclyl and
bis-oxy-C.sub.1-3 alkylene. The other variables are as defined in
the claims. The compounds are useful for the treatment of
proliferative diseases. ##STR00001##
Inventors: |
Howard; Philip Wilson (London,
GB), Gregson; Stephen John (London, GB),
Masterson; Luke (London, GB) |
Applicant: |
Name |
City |
State |
Country |
Type |
Howard; Philip Wilson
Gregson; Stephen John
Masterson; Luke |
London
London
London |
N/A
N/A
N/A |
GB
GB
GB |
|
|
Assignee: |
Spirogen Sarl (St-Legier-la
Chiesaz, CH)
|
Family
ID: |
40097621 |
Appl.
No.: |
13/123,327 |
Filed: |
October 16, 2009 |
PCT
Filed: |
October 16, 2009 |
PCT No.: |
PCT/GB2009/002498 |
371(c)(1),(2),(4) Date: |
April 08, 2011 |
PCT
Pub. No.: |
WO2010/043880 |
PCT
Pub. Date: |
April 22, 2010 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20110196148 A1 |
Aug 11, 2011 |
|
Foreign Application Priority Data
|
|
|
|
|
Oct 17, 2008 [GB] |
|
|
0819095.1 |
|
Current U.S.
Class: |
540/496 |
Current CPC
Class: |
A61P
35/02 (20180101); A61P 35/00 (20180101); A61P
43/00 (20180101); C07D 487/04 (20130101) |
Current International
Class: |
C07D
487/04 (20060101) |
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|
Primary Examiner: Loewe; Sun Jae
Attorney, Agent or Firm: Michael Best & Friedrich
LLP
Claims
The invention claimed is:
1. A compound with the formula I: ##STR00034## wherein: R.sup.2 is
of formula II: ##STR00035## where A is a C.sub.5-7 aryl group, X is
selected from the group consisting of: OH, SH, CO.sub.2H, COH,
N.dbd.C.dbd.O, NHR.sup.N, wherein R.sup.N is selected from the
group consisting of H and C.sub.1-4 alkyl, and
(OC.sub.2H.sub.4).sub.mOCH.sub.3, where m is 1 to 3, and either:
(i) Q.sup.1 is a single bond, and Q.sup.2 is selected from the
group consisting of a single bond and --Z--(CH.sub.2).sub.n--,
where Z is selected from the group consisting of a single bond, O,
S and NH and n is from 1 to 3; or (ii) Q.sup.1 is --CH.dbd.CH--,
and Q.sup.2 is a single bond; R.sup.12 is a C.sub.5-10 aryl group,
optionally substituted by one or more substituents selected from
the group consisting of: halo, nitro, cyano, ether, C.sub.1-7
alkyl, C.sub.3-7 heterocyclyl and bis-oxy-C.sub.1-3 alkylene;
R.sup.6 and R.sup.9 are independently selected from the group
consisting of H, R, OH, OR, SH, SR, NH.sub.2, NHR, NRR', nitro,
Me.sub.3Sn and halo; where R and R' are independently selected from
the group consisting of optionally substituted C.sub.1-12 alkyl,
C.sub.3-20 heterocyclyl and C.sub.5-20 aryl groups; R.sup.7 is
selected from the group consisting of H, R, OH, OR, SH, SR,
NH.sub.2, NHR, NHRR', nitro, Me.sub.3Sn and halo; either: (a)
R.sup.10 is H, and R.sup.11 is OH, or OR.sup.A, where R.sup.A is
C.sub.1-4 alkyl; (b) R.sup.10 and R.sup.11 form a nitrogen-carbon
double bond between the nitrogen and carbon atoms to which they are
bound; or (c) R.sup.10 is H and R.sup.11 is SO.sub.zM, where z is 2
or 3 and M is a monovalent pharmaceutically acceptable cation; R''
is a C.sub.3-12 alkylene group, which chain may be interrupted by
one or more heteroatoms selected from the group consisting of O, S,
and NH, and/or aromatic rings selected from benzene or pyridine; Y
and Y' are independently selected from the group consisting of O,
S, or NH; R.sup.6', R.sup.7', R.sup.9' are selected from the same
groups as R.sup.6, R.sup.7 and R.sup.9 respectively and R.sup.10'
and R.sup.11' are the same as R.sup.10 and R.sup.11, wherein if
R.sup.11 and R.sup.11' are SO.sub.zM, M represents a divalent
pharmaceutically acceptable cation selected from Ca.sup.2+ and
Mg.sup.2+.
2. A compound according to claim 1, wherein R.sup.7 is selected
from H, OH and OR.
3. A compound according to claim 2, wherein R.sup.7 is a C.sub.1-4
alkyloxy group.
4. A compound according to claim 1, wherein Y is O.
5. A compound according to claim 1, wherein R'' is C.sub.3-7
alkylene.
6. A compound according to claim 1, wherein R.sup.9 is H.
7. A compound according to claim 1, wherein R.sup.6 is selected
from H and halo.
8. A compound according to claim 1, wherein A is phenyl.
9. A compound according to claim 1, wherein X is selected from OH,
SH, or NH.sub.2.
10. A compound according to claim 1, wherein Q.sup.1 is a single
bond.
11. A compound according to claim 10, wherein Q.sup.2 is a single
bond.
12. A compound according to claim 10, wherein Q.sup.2 is
--Z--(CH.sub.2).sub.n--, Z is O or S and n is 1 or 2.
13. A compound according to claim 1, wherein Q.sup.1 is
--CH.dbd.CH--.
14. A compound according to claim 1, wherein R.sup.12 is a
C.sub.5-7 aryl group.
15. A compound according to claim 14, wherein R.sup.12 is
phenyl.
16. A compound according to claim 1, wherein R.sup.12 is a
C.sub.8-10 aryl group.
17. A compound according to claim 1, wherein R.sup.12 bears one to
three substituent groups.
18. A compound according to claim 1, wherein R.sup.10 and R.sup.11
and form a nitrogen-carbon double bond.
19. A compound according to claim 1, wherein R.sup.6', R.sup.7',
R.sup.9', R.sup.10', R.sup.11' and Y' are the same as R.sup.6,
R.sup.7, R.sup.9, R.sup.10, R.sup.11 and Y respectively.
20. A compound of formula II: ##STR00036## wherein: R.sup.2 is
##STR00037## where A is a C.sub.5-7 aryl group, X is selected from
the group consisting of: OH, SH, CO.sub.2H, COH, N.dbd.C.dbd.O,
NHR.sup.N, wherein R.sup.N is selected from the group consisting of
H and C.sub.1-4 alkyl, and (OC.sub.2H.sub.4).sub.mOCH.sub.3, where
m is 1 to 3, and either: (i) Q.sup.1 is a single bond, and Q.sup.2
is selected from a single bond or --Z--(CH.sub.2).sub.n--, where Z
is selected from a single bond, O, S or NH and is from 1 to 3; or
(ii) Q.sup.1 is --CH.dbd.CH--, and Q.sup.2 is a single bond;
R.sup.12 is a C.sub.5-10 aryl group, optionally substituted by one
or more substituents selected from the group consisting of: halo,
nitro, cyano, ether, C.sub.1-7 alkyl, C.sub.3-7 heterocyclyl and
bis-oxy-C.sub.1-3 alkylene; R.sup.6 and R.sup.9 are independently
selected from the group consisting of H, R, OH, OR, SH, SR,
NH.sub.2, NHR, NRR', nitro, Me.sub.3Sn and halo; where R and R' are
independently selected from the group consisting of optionally
substituted C.sub.1-12 alkyl, C.sub.3-20 heterocyclyl and
C.sub.5-20 aryl groups; R.sup.7 is selected from the group
consisting of H, R, OH, OR, SH, SR, NH.sub.2, NHR, NHRR', nitro,
Me.sub.3Sn and halo; either: (a) R.sup.10 is carbamate nitrogen
protecting group, and R.sup.11 is O-Prot.sup.o, wherein Prot.sup.o
is an oxygen protecting group; (b) R.sup.10 is a hemi-aminal
nitrogen protecting group and R.sup.11 is an oxo group; R'' is a
C.sub.3-12 alkylene group, which chain may be interrupted by one or
more heteroatoms, selected from the group consisting of O, S, and
NR.sup.N2 (where R.sup.N2 is H or C.sub.1-4 alkyl), and/or aromatic
rings, e.g. benzene or pyridine; Y and Y' are selected from O, S,
or NH; R.sup.6', R.sup.7', R.sup.9' are selected from the same
groups as R.sup.6, R.sup.7 and R.sup.9 respectively and R.sup.10'
and R.sup.11' are the same as R.sup.10 and R.sup.11.
21. A compound according to claim 20, wherein R.sup.7 is selected
from H, OH or OR.
22. A compound according to claim 21, wherein R.sup.7 is a
C.sub.1-4 alkyloxy group.
23. A compound according to claim 20, wherein Y is O.
24. A compound according to claim 20, wherein R'' is C.sub.3-7
alkylene.
25. A compound according to claim 20, wherein R.sup.9 is H.
26. A compound according to claim 20, wherein R.sup.6 is selected
from H or halo.
27. A compound according to claim 20, wherein A is phenyl.
28. A compound according to claim 20, wherein X is selected from
OH, SH, or NH.sub.2.
29. A compound according to claim 20, wherein Q.sup.1 is a single
bond.
30. A compound according to claim 29, wherein Q.sup.2 is a single
bond.
31. A compound according to claim 29, wherein Q.sup.2 is
--Z--(CH.sub.2).sub.n--, Z is O or S and n is 1 or 2.
32. A compound according to claim 20, wherein Q.sup.1 is
--CH.dbd.CH--.
33. A compound according to claim 20, wherein R.sup.12 is a
C.sub.5-7 aryl group.
34. A compound according to claim 33, wherein R.sup.12 is
phenyl.
35. A compound according to claim 20, wherein R.sup.12 is a
C.sub.8-10 aryl group.
36. A compound according to claim 20, wherein R.sup.12 bears one to
three substituent groups.
37. A compound according to claim 20, wherein R.sup.10 is Troc.
38. A compound according to claim 20, wherein R.sup.11 is OTBS.
39. A compound according to claim 20, wherein R.sup.11 is oxo and
R.sup.10 is SEM.
40. A compound according to claim 20, wherein R.sup.6', R.sup.7',
R.sup.9', R.sup.10', R.sup.11' and Y' are the same as R.sup.6,
R.sup.7, R.sup.9, R.sup.10, R.sup.11 and Y respectively.
41. A compound of the formula: ##STR00038##
42. A compound of the formula: ##STR00039##
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
This patent application is a national stage filing under 35 U.S.C.
371 of International Application No. PCT/GB2009/002498, filed on
Oct. 16, 2009, which claims foreign priority benefits to United
Kingdom Patent Application No. 0819095.1, filed Oct. 17, 2008.
These applications are incorporated herein by reference in their
entireties.
The present invention relates to pyrrolobenzodiazepines (PBDs), in
particular pyrrolobenzodiazepine dimers having a C2-C3 double bond
and an aryl group at the C2 position in each monomer unit.
BACKGROUND TO THE INVENTION
Some pyrrolobenzodiazepines (PBDs) have the ability to recognise
and bond to specific sequences of DNA; the preferred sequence is
PuGPu. The first PBD antitumour antibiotic, anthramycin, was
discovered in 1965 (Leimgruber, et al., J. Am. Chem. Soc., 87,
5793-5795 (1965); Leimgruber, et al., J. Am. Chem. Soc., 87,
5791-5793 (1965)). Since then, a number of naturally occurring PBDs
have been reported, and over 10 synthetic routes have been
developed to a variety of analogues (Thurston, et al., Chem. Rev.
1994, 433-465 (1994)). Family members include abbeymycin
(Hochlowski, et al., J. Antibiotics, 40, 145-148 (1987)),
chicamycin (Konishi, et al., J. Antibiotics, 37, 200-206 (1984)),
DC-81 (Japanese Patent 58-180 487; Thurston, et al., Chem. Brit.,
26, 767-772 (1990); Bose, et al., Tetrahedron, 48, 751-758 (1992)),
mazethramycin (Kuminoto, et al., J. Antibiotics, 33, 665-667
(1980)), neothramycins A and B (Takeuchi, et al., J. Antibiotics,
29, 93-96 (1976)), porothramycin (Tsunakawa, et al., J.
Antibiotics, 41, 1366-1373 (1988)), prothracarcin (Shimizu, et al,
J. Antibiotics, 29, 2492-2503 (1982); Langley and Thurston, J. Org.
Chem., 52, 91-97 (1987)), sibanomicin (DC-102)(Hara, et al., J.
Antibiotics, 41, 702-704 (1988); Itoh, et al., J. Antibiotics, 41,
1281-1284 (1988)), sibiromycin (Leber, et al., J. Am. Chem. Soc.,
110, 2992-2993 (1988)) and tomamycin (Arima, et al., J.
Antibiotics, 25, 437-444 (1972)). PBDs are of the general
structure:
##STR00002##
They differ in the number, type and position of substituents, in
both their aromatic A rings and pyrrolo C rings, and in the degree
of saturation of the C ring. In the B-ring there is either an imine
(N.dbd.C), a carbinolamine(NH--CH(OH)), or a carbinolamine methyl
ether (NH--CH(OMe)) at the N10-C11 position which is the
electrophilic centre responsible for alkylating DNA. All of the
known natural products have an (S)-configuration at the chiral C11a
position which provides them with a right-handed twist when viewed
from the C ring towards the A ring. This gives them the appropriate
three-dimensional shape for isohelicity with the minor groove of
B-form DNA, leading to a snug fit at the binding site (Kohn, In
Antibiotics III. Springer-Verlag, New York, pp. 3-11 (1975); Hurley
and Needham-VanDevanter, Acc. Chem. Res., 19, 230-237 (1986)).
Their ability to form an adduct in the minor groove, enables them
to interfere with DNA processing, hence their use as antitumour
agents.
It has been previously disclosed that the biological activity of
this molecules can be potentiated by joining two PBD units together
through their C8/C'-hydroxyl functionalities via a flexible
alkylene linker (Bose, D. S., et al., J. Am. Chem. Soc., 114,
4939-4941 (1992); Thurston, D. E., et al., J. Org. Chem., 61,
8141-8147 (1996)). The PBD dimers are thought to form
sequence-selective DNA lesions such as the palindromic
5'-Pu-GATC-Py-3' interstrand cross-link (Smellie, M., et al.,
Biochemistry, 42, 8232-8239 (2003); Martin, C., et al.,
Biochemistry, 44, 4135-4147) which is thought to be mainly
responsible for their biological activity. One example of a PBD
dimmer, SG2000 (SJG-136):
##STR00003## has recently completed Phase I clinical trials in the
oncology area and is about to enter Phase II (Gregson, S., et al.,
J. Med. Chem., 44, 737-748 (2001); Alley, M. C., et al., Cancer
Research, 64, 6700-6706 (2004); Hartley, J. A., et al., Cancer
Research, 64, 6693-6699 (2004)).
More recently, the present inventors have previously disclosed in
WO 2005/085251, dimeric PBD compounds bearing C2 aryl substituents,
such as SG2202 (ZC-207):
##STR00004## and in WO2006/111759, bisulphites of such PBD
compounds, for example SG2285 (ZC-423):
##STR00005##
These compounds have been shown to be highly useful cytotoxic
agents (Howard, P. W., et al., Bioorg. Med. Chem. (2009), doi:
10.1016/j.bmcl.2009.09.012).
Due to the manner in which these highly potent compounds act in
cross-linking DNA, these molecules have been made symmetrically.
This provides for straightforward synthesis, either by constructing
the PBD moieties simultaneously having already formed the dimer
linkage, or by reacting already constructed PBD moieties with the
dimer linking group.
DISCLOSURE OF THE INVENTION
The present inventors have developed an unsymmetrical dimeric PBD
compound bearing aryl groups in the C2 position of each monomer,
where one of these groups bears a substituent designed to provide
an anchor for linking the compound to another moiety.
The present invention comprises a compound with the formula I:
##STR00006## wherein: R.sup.2 is of formula II:
##STR00007## where A is a C.sub.5-7 aryl group, X is selected from
the group comprising: OH, SH, CO.sub.2H, COH, N.dbd.C.dbd.O,
NHR.sup.N, wherein R.sup.N is selected from the group comprising H
and C.sub.1-4 alkyl, and (OC.sub.2H.sub.4).sub.mOCH.sub.3, where m
is 1 to 3, and either: (i) Q.sup.1 is a single bond, and Q.sup.2 is
selected from a single bond and --Z--(CH.sub.2).sub.n, where Z is
selected from a single bond, O, S and NH and is from 1 to 3; or
(ii) Q.sup.1 is --CH.dbd.CH--, and Q.sup.2 is a single bond;
R.sup.12 is a C.sub.5-10 aryl group, optionally substituted by one
or more substituents selected from the group comprising: halo,
nitro, cyano, ether, C.sub.1-7 alkyl, C.sub.3-7 heterocyclyl and
bis-oxy-C.sub.1-3 alkylene; R.sup.6 and R.sup.9 are independently
selected from H, R, OH, OR, SH, SR, NH.sub.2, NHR, NRR', nitro,
Me.sub.3Sn and halo; where R and R' are independently selected from
optionally substituted C.sub.1-12 alkyl, C.sub.3-20 heterocyclyl
and C.sub.5-20 aryl groups; R.sup.7 is selected from H, R, OH, OR,
SH, SR, NH.sub.2, NHR, NHRR', nitro, Me.sub.3Sn and halo; either:
(a) R.sup.19 is H, and R.sup.11 is OH, OR.sup.A, where R.sup.A is
C.sub.1-4 alkyl; (b) R.sup.19 and R.sup.11 form a nitrogen-carbon
double bond between the nitrogen and carbon atoms to which they are
bound; or (c) R.sup.19 is H and R.sup.11 is SO.sub.zM, where z is 2
or 3 and M is a monovalent pharmaceutically acceptable cation; R''
is a C.sub.3-12 alkylene group, which chain may be interrupted by
one or more heteroatoms, e.g. O, S, NR.sup.N2 (where R.sup.N2 is H
or C.sub.1-4 alkyl), and/or aromatic rings, e.g. benzene or
pyridine; Y and Y' are selected from O, S, or NH; R.sup.6',
R.sup.7', R.sup.9' are selected from the same groups as R.sup.6,
R.sup.7 and R.sup.9 respectively and R.sup.10' and R.sup.11' are
the same as R.sup.10 and R.sup.11, wherein if R.sup.11 and
R.sup.11' are SO.sub.zM, M may represent a divalent
pharmaceutically acceptable cation.
A second aspect of the present invention provides the use of a
compound of the first aspect of the invention in the manufacture of
a medicament for treating a proliferative disease. The second
aspect also provides a compound of the first aspect of the
invention for use in the treatment of a proliferative disease.
One of ordinary skill in the art is readily able to determine
whether or not a candidate conjugate treats a proliferative
condition for any particular cell type. For example, assays which
may conveniently be used to assess the activity offered by a
particular compound are described in the examples below.
The term "proliferative disease" pertains to an unwanted or
uncontrolled cellular proliferation of excessive or abnormal cells
which is undesired, such as, neoplastic or hyperplastic growth,
whether in vitro or in vivo.
Examples of proliferative conditions include, but are not limited
to, benign, pre-malignant, and malignant cellular proliferation,
including but not limited to, neoplasms and tumours (e.g.
histocytoma, glioma, astrocyoma, osteoma), cancers (e.g. lung
cancer, small cell lung cancer, gastrointestinal cancer, bowel
cancer, colon cancer, breast carinoma, ovarian carcinoma, prostate
cancer, testicular cancer, liver cancer, kidney cancer, bladder
cancer, pancreas cancer, brain cancer, sarcoma, osteosarcoma,
Kaposi's sarcoma, melanoma), leukemias, psoriasis, bone diseases,
fibroproliferative disorders (e.g. of connective tissues), and
atherosclerosis. Cancers of particular interest include, but are
not limited to, leukemias and ovarian cancers.
Any type of cell may be treated, including but not limited to,
lung, gastrointestinal (including, e.g. bowel, colon), breast
(mammary), ovarian, prostate, liver (hepatic), kidney (renal),
bladder, pancreas, brain, and skin.
A third aspect of the present invention comprises a compound of
formula II:
##STR00008## wherein: R.sup.2 is of formula II.
##STR00009## where A is a C.sub.5-7 aryl group, X is selected from
the group comprising: OH, SH, CO.sub.2H, COH, N.dbd.C.dbd.O,
NHR.sup.N, wherein R.sup.N is selected from the group comprising H
and C.sub.1-4 alkyl, and (OC.sub.2H.sub.4).sub.mOCH.sub.3, where m
is 1 to 3, and either: (i) Q.sup.1 is a single bond, and Q.sup.2 is
selected from a single bond and --Z--(CH.sub.2).sub.n--, where Z is
selected from a single bond, O, S and NH and is from 1 to 3; or
(ii) Q.sup.1 is --CH.dbd.CH--, and Q.sup.2 is a single bond;
R.sup.12 is a C.sub.5-10 aryl group, optionally substituted by one
or more substituents selected from the group comprising: halo,
nitro, cyano, ether, C.sub.1-7 alkyl, C.sub.3-7 heterocyclyl and
bis-oxy-C.sub.1-3 alkylene; R.sup.6 and R.sup.9 are independently
selected from H, R, OH, OR, SH, SR, NH.sub.2, NHR, NRR', nitro,
Me.sub.3Sn and halo; where R and R' are independently selected from
optionally substituted C.sub.1-12 alkyl, C.sub.3-20 heterocyclyl
and C.sub.5-20 aryl groups; R.sup.7 is selected from H, R, OH, OR,
SH, SR, NH.sub.2, NHR, NHRR', nitro, Me.sub.3Sn and halo; either:
(a) R.sup.19 is carbamate nitrogen protecting group, and R.sup.11
is O-Prot.sup.o, wherein Prot.sup.o is an oxygen protecting group;
(b) R.sup.19 is a hemi-aminal nitrogen protecting group and
R.sup.11 is an oxo group; R'' is a C.sub.3-12 alkylene group, which
chain may be interrupted by one or more heteroatoms, e.g. O, S,
NR.sup.N2 (where R.sup.N2 is H or C.sub.1-4 alkyl), and/or aromatic
rings, e.g. benzene or pyridine; Y and Y' are selected from O, S,
or NH; R.sup.6', R.sup.7', R.sup.9' are selected from the same
groups as R.sup.6, R.sup.7 and R.sup.9 respectively and R.sup.10'
and R.sup.11' are the same as R.sup.19 and R.sup.11.
A fourth aspect of the present invention comprises a method of
making a compound of formula I from a compound of formula II by
deprotection of the imine bond.
The unsymmetrical dimeric PBD compounds of the present invention
are made by different strategies to those previously employed in
making symmetrical dimeric PBD compounds. In particular, the
present inventors have developed a method which involves adding
each each C2 aryl substituent to a symmetrical PBD dimer core in
separate method steps. Accordingly, a fifth aspect of the present
invention provides a method of making a compound of the first or
third aspect of the invention, comprising at least one of the
method steps set out below.
DEFINITIONS
Pharmaceutically Acceptable Cations
Examples of pharmaceutically acceptable monovalent and divalent
cations are discussed in Berge, et al., J. Pharm. Sci., 66, 1-19
(1977), which is incorporated herein by reference.
The pharmaceutically acceptable cation may be inorganic or
organic.
Examples of pharmaceutically acceptable monovalent inorganic
cations include, but are not limited to, alkali metal ions such as
Na.sup.+ and K.sup.+. Examples of pharmaceutically acceptable
divalent inorganic cations include, but are not limited to,
alkaline earth cations such as Ca.sup.2+ and Mg.sup.2+. Examples of
pharmaceutically acceptable organic cations include, but are not
limited to, ammonium ion (i.e. NH.sub.4.sup.+) and substituted
ammonium ions (e.g. NH.sub.3R.sup.+, NH.sub.2R.sub.2.sup.+,
NHR.sub.3.sup.+, NR.sub.4.sup.+). Examples of some suitable
substituted ammonium ions are those derived from: ethylamine,
diethylamine, dicyclohexylamine, triethylamine, butylamine,
ethylenediamine, ethanolamine, diethanolamine, piperazine,
benzylamine, phenylbenzylamine, choline, meglumine, and
tromethamine, as well as amino acids, such as lysine and arginine.
An example of a common quaternary ammonium ion is
N(CH.sub.3).sub.4.sup.+.
Substituents
The phrase "optionally substituted" as used herein, pertains to a
parent group which may be unsubstituted or which may be
substituted.
Unless otherwise specified, the term "substituted" as used herein,
pertains to a parent group which bears one or more substituents.
The term "substituent" is used herein in the conventional sense and
refers to a chemical moiety which is covalently attached to, or if
appropriate, fused to, a parent group. A wide variety of
substituents are well known, and methods for their formation and
introduction into a variety of parent groups are also well
known.
Examples of substituents are described in more detail below.
C.sub.1-12 alkyl: The term "C.sub.1-12 alkyl" as used herein,
pertains to a monovalent moiety obtained by removing a hydrogen
atom from a carbon atom of a hydrocarbon compound having from 1 to
12 carbon atoms, which may be aliphatic or alicyclic, and which may
be saturated or unsaturated (e.g. partially unsaturated, fully
unsaturated). Thus, the term "alkyl" includes the sub-classes
alkenyl, alkynyl, cycloalkyl, etc., discussed below.
Examples of saturated alkyl groups include, but are not limited to,
methyl (C.sub.1), ethyl (C.sub.2), propyl (C.sub.3), butyl
(C.sub.4), pentyl (C.sub.5), hexyl (C.sub.6) and heptyl
(C.sub.7).
Examples of saturated linear alkyl groups include, but are not
limited to, methyl (C.sub.1), ethyl (C.sub.2), n-propyl (C.sub.3),
n-butyl (C.sub.4), n-pentyl (amyl) (C.sub.5), n-hexyl (C.sub.6) and
n-heptyl (C.sub.7).
Examples of saturated branched alkyl groups include iso-propyl
(C.sub.3), iso-butyl (C.sub.4), sec-butyl (C.sub.4), tert-butyl
(C.sub.4), iso-pentyl (C.sub.5), and neo-pentyl (C.sub.5).
C.sub.2-12 Alkenyl: The term "C.sub.2-12 alkenyl" as used herein,
pertains to an alkyl group having one or more carbon-carbon double
bonds.
Examples of unsaturated alkenyl groups include, but are not limited
to, ethenyl (vinyl, --CH.dbd.CH.sub.2), 1-propenyl
(--CH.dbd.CH--CH.sub.3), 2-propenyl (allyl, --CH--CH.dbd.CH.sub.2),
isopropenyl (1-methylvinyl, --C(CH.sub.3).dbd.CH.sub.2), butenyl
(C.sub.4), pentenyl (C.sub.5), and hexenyl (C.sub.6).
C.sub.2-12 alkynyl: The term "C.sub.2-12 alkynyl" as used herein,
pertains to an alkyl group having one or more carbon-carbon triple
bonds.
Examples of unsaturated alkynyl groups include, but are not limited
to, ethynyl (--C.ident.CH) and 2-propynyl (propargyl,
--CH.sub.2--C.ident.CH).
C.sub.3-12 cycloalkyl: The term "C.sub.3-12 cycloalkyl" as used
herein, pertains to an alkyl group which is also a cyclyl group;
that is, a monovalent moiety obtained by removing a hydrogen atom
from an alicyclic ring atom of a cyclic hydrocarbon (carbocyclic)
compound, which moiety has from 3 to 7 carbon atoms, including from
3 to 7 ring atoms.
Examples of cycloalkyl groups include, but are not limited to,
those derived from: saturated monocyclic hydrocarbon compounds:
cyclopropane (C.sub.3), cyclobutane (C.sub.4), cyclopentane
(C.sub.5), cyclohexane (C.sub.6), cycloheptane (C.sub.7),
methylcyclopropane (C.sub.4), dimethylcyclopropane (C.sub.5),
methylcyclobutane (C.sub.5), dimethylcyclobutane (C.sub.6),
methylcyclopentane (C.sub.6), dimethylcyclopentane (C.sub.7) and
methylcyclohexane (C.sub.7); unsaturated monocyclic hydrocarbon
compounds: cyclopropene (C.sub.3), cyclobutene (C.sub.4),
cyclopentene (C.sub.5), cyclohexene (C.sub.6), methylcyclopropene
(C.sub.4), dimethylcyclopropene (C.sub.5), methylcyclobutene
(C.sub.5), dimethylcyclobutene (C.sub.6), methylcyclopentene
(C.sub.6), dimethylcyclopentene (C.sub.7) and methylcyclohexene
(C.sub.7); and saturated polycyclic hydrocarbon compounds:
norcarane (C.sub.7), norpinane (C.sub.7), norbornane (C.sub.7).
C.sub.3-20 heterocyclyl: The term "C.sub.3-20 heterocyclyl" as used
herein, pertains to a monovalent moiety obtained by removing a
hydrogen atom from a ring atom of a heterocyclic compound, which
moiety has from 3 to 20 ring atoms, of which from 1 to 10 are ring
heteroatoms. Preferably, each ring has from 3 to 7 ring atoms, of
which from 1 to 4 are ring heteroatoms.
In this context, the prefixes (e.g. C.sub.3-20, C.sub.3-7,
C.sub.5-6, etc.) denote the number of ring atoms, or range of
number of ring atoms, whether carbon atoms or heteroatoms. For
example, the term "C.sub.5-6heterocyclyl", as used herein, pertains
to a heterocyclyl group having 5 or 6 ring atoms.
Examples of monocyclic heterocyclyl groups include, but are not
limited to, those derived from:
N.sub.1: aziridine (C.sub.3), azetidine (C.sub.4), pyrrolidine
(tetrahydropyrrole) (C.sub.5), pyrroline (e.g., 3-pyrroline,
2,5-dihydropyrrole) (C.sub.5), 2H-pyrrole or 3H-pyrrole
(isopyrrole, isoazole) (C.sub.5), piperidine (C.sub.6),
dihydropyridine (C.sub.6), tetrahydropyridine (C.sub.6), azepine
(C.sub.7); O.sub.1: oxirane (C.sub.3), oxetane (C.sub.4), oxolane
(tetrahydrofuran) (C.sub.5), oxole (dihydrofuran) (C.sub.5), oxane
(tetrahydropyran) (C.sub.6), dihydropyran (C.sub.6), pyran
(C.sub.6), oxepin (C.sub.7); S.sub.1: thiirane (C.sub.3), thietane
(C.sub.4), thiolane (tetrahydrothiophene) (C.sub.5), thiane
(tetrahydrothiopyran) (C.sub.6), thiepane (C.sub.7); O.sub.2:
dioxolane (C.sub.5), dioxane (C.sub.6), and dioxepane (C.sub.7);
O.sub.3: trioxane (C.sub.6); N.sub.2: imidazolidine (C.sub.5),
pyrazolidine (diazolidine) (C.sub.5), imidazoline (C.sub.5),
pyrazoline (dihydropyrazole) (C.sub.5), piperazine (C.sub.6);
N.sub.1O.sub.1: tetrahydrooxazole (C.sub.5), dihydrooxazole
(C.sub.5), tetrahydroisoxazole (C.sub.5), dihydroisoxazole
(C.sub.5), morpholine (C.sub.6), tetrahydrooxazine (C.sub.6),
dihydrooxazine (C.sub.6), oxazine (C.sub.6); N.sub.1S.sub.1:
thiazoline (C.sub.5), thiazolidine (C.sub.5), thiomorpholine
(C.sub.6); N.sub.2O.sub.1: oxadiazine (C.sub.6); O.sub.1S.sub.1:
oxathiole (C.sub.5) and oxathiane (thioxane) (C.sub.6); and,
N.sub.1O.sub.1S.sub.1: oxathiazine (C.sub.6).
Examples of substituted monocyclic heterocyclyl groups include
those derived from saccharides, in cyclic form, for example,
furanoses (C.sub.5), such as arabinofuranose, lyxofuranose,
ribofuranose, and xylofuranse, and pyranoses (C.sub.6), such as
allopyranose, altropyranose, glucopyranose, mannopyranose,
gulopyranose, idopyranose, galactopyranose, and talopyranose.
C.sub.5-20 aryl: The term "C.sub.5-20 aryl", as used herein,
pertains to a monovalent moiety obtained by removing a hydrogen
atom from an aromatic ring atom of an aromatic compound, which
moiety has from 3 to 20 ring atoms. Preferably, each ring has from
5 to 7 ring atoms.
In this context, the prefixes (e.g. C.sub.3-20, C.sub.5-7,
C.sub.5-6, etc.) denote the number of ring atoms, or range of
number of ring atoms, whether carbon atoms or heteroatoms. For
example, the term "C.sub.5-6 aryl" as used herein, pertains to an
aryl group having 5 or 6 ring atoms.
The ring atoms may be all carbon atoms, as in "carboaryl
groups".
Examples of carboaryl groups include, but are not limited to, those
derived from benzene (i.e. phenyl) (C.sub.6), naphthalene
(C.sub.10), azulene (C.sub.10), anthracene (C.sub.14), phenanthrene
(C.sub.14), naphthacene (C.sub.18), and pyrene (C.sub.16).
Examples of aryl groups which comprise fused rings, at least one of
which is an aromatic ring, include, but are not limited to, groups
derived from indane (e.g. 2,3-dihydro-1H-indene) (C.sub.9), indene
(C.sub.9), isoindene (C.sub.9), tetraline
(1,2,3,4-tetrahydronaphthalene (C.sub.10), acenaphthene (C.sub.12),
fluorene (C.sub.13), phenalene (C.sub.13), acephenanthrene
(C.sub.15), and aceanthrene (C.sub.16).
Alternatively, the ring atoms may include one or more heteroatoms,
as in "heteroaryl groups". Examples of monocyclic heteroaryl groups
include, but are not limited to, those derived from:
N.sub.1: pyrrole (azole) (C.sub.5), pyridine (azine) (C.sub.6);
O.sub.1: furan (oxole) (C.sub.5);
S.sub.1: thiophene (thiole) (C.sub.5);
N.sub.1O.sub.1: oxazole (C.sub.5), isoxazole (C.sub.5), isoxazine
(C.sub.6);
N.sub.2O.sub.1: oxadiazole (furazan) (C.sub.5);
N.sub.3O.sub.1: oxatriazole (C.sub.5);
N.sub.1S.sub.1: thiazole (C.sub.5), isothiazole (C.sub.5);
N.sub.2: imidazole (1,3-diazole) (C.sub.5), pyrazole (1,2-diazole)
(C.sub.5), pyridazine (1,2-diazine) (C.sub.6), pyrimidine
(1,3-diazine) (C.sub.6) (e.g., cytosine, thymine, uracil), pyrazine
(1,4-diazine) (C.sub.6);
N.sub.3: triazole (C.sub.5), triazine (C.sub.6); and,
N.sub.4: tetrazole (C.sub.5).
Examples of heteroaryl which comprise fused rings, include, but are
not limited to: C.sub.9 (with 2 fused rings) derived from
benzofuran (O.sub.1), isobenzofuran (O.sub.1), indole (N.sub.1),
isoindole (N.sub.1), indolizine (N.sub.1), indoline (N.sub.1),
isoindoline (N.sub.1), purine (N.sub.4) (e.g., adenine, guanine),
benzimidazole (N.sub.2), indazole (N.sub.2), benzoxazole
(N.sub.1O.sub.1), benzisoxazole (N.sub.1O.sub.1), benzodioxole
(O.sub.2), benzofurazan (N.sub.2O.sub.1), benzotriazole (N.sub.3),
benzothiofuran (S.sub.1), benzothiazole (N.sub.1S.sub.1),
benzothiadiazole (N.sub.2S); C.sub.10 (with 2 fused rings) derived
from chromene (O.sub.1), isochromene (O.sub.1), chroman (O.sub.1),
isochroman (O.sub.1), benzodioxan (O.sub.2), quinoline (N.sub.1),
isoquinoline (N.sub.1), quinolizine (N.sub.1), benzoxazine
(N.sub.1O.sub.1), benzodiazine (N.sub.2), pyridopyridine (N.sub.2),
quinoxaline (N.sub.2), quinazoline (N2), cinnoline (N.sub.2),
phthalazine (N.sub.2), naphthyridine (N.sub.2), pteridine
(N.sub.4); C.sub.11 (with 2 fused rings) derived from
benzodiazepine (N.sub.2); C.sub.13 (with 3 fused rings) derived
from carbazole (N.sub.1), dibenzofuran (O.sub.1), dibenzothiophene
(S.sub.1), carboline (N.sub.2), perimidine (N.sub.2), pyridoindole
(N.sub.2); and, C.sub.14 (with 3 fused rings) derived from acridine
(N.sub.1), xanthene (O.sub.1), thioxanthene (S.sub.1), oxanthrene
(O.sub.2), phenoxathiin (O.sub.1S.sub.1), phenazine (N.sub.2),
phenoxazine (N.sub.1O.sub.1), phenothiazine (N.sub.1S.sub.1),
thianthrene (S.sub.2), phenanthridine (N.sub.1), phenanthroline
(N.sub.2), phenazine (N.sub.2).
The above groups, whether alone or part of another substituent, may
themselves optionally be substituted with one or more groups
selected from themselves and the additional substituents listed
below.
Halo: --F, --Cl, --Br, and --I.
Hydroxy: --OH.
Ether: --OR, wherein R is an ether substituent, for example, a
C.sub.1-7 alkyl group (also referred to as a C.sub.1-7 alkoxy
group, discussed below), a C.sub.3-20 heterocyclyl group (also
referred to as a C.sub.3-20 heterocyclyloxy group), or a C.sub.5-20
aryl group (also referred to as a C.sub.5-20 aryloxy group),
preferably a C.sub.1-7alkyl group.
Alkoxy: --OR, wherein R is an alkyl group, for example, a C.sub.1-7
alkyl group. Examples of C.sub.1-7 alkoxy groups include, but are
not limited to, --OMe (methoxy), --OEt (ethoxy), --O(nPr)
(n-propoxy), --O(iPr) (isopropoxy), --O(nBu) (n-butoxy), --O(sBu)
(sec-butoxy), --O(iBu) (isobutoxy), and --O(tBu) (tert-butoxy).
Acetal: --CH(OR.sup.1)(OR.sup.2), wherein R.sup.1 and R.sup.2 are
independently acetal substituents, for example, a C.sub.1-7 alkyl
group, a C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group, or, in the case of a "cyclic"
acetal group, R.sup.1 and R.sup.2, taken together with the two
oxygen atoms to which they are attached, and the carbon atoms to
which they are attached, form a heterocyclic ring having from 4 to
8 ring atoms. Examples of acetal groups include, but are not
limited to, --CH(OMe).sub.2, --CH(OEt).sub.2, and
--CH(OMe)(OEt).
Hemiacetal: --CH(OH)(OR.sup.1), wherein R.sup.1 is a hemiacetal
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of hemiacetal groups include, but
are not limited to, --CH(OH)(OMe) and --CH(OH)(OEt).
Ketal: --CR(OR.sup.1)(OR.sup.2), where R.sup.1 and R.sup.2 are as
defined for acetals, and R is a ketal substituent other than
hydrogen, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples ketal groups include, but are not
limited to, --C(Me)(OMe).sub.2, --C(Me)(OEt).sub.2,
--C(Me)(OMe)(OEt), --C(Et)(OMe).sub.2, --C(Et)(OEt).sub.2, and
--C(Et)(OMe)(OEt).
Hemiketal: --CR(OH)(OR.sup.1), where R.sup.1 is as defined for
hemiacetals, and R is a hemiketal substituent other than hydrogen,
for example, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl
group. Examples of hemiacetal groups include, but are not limited
to, --C(Me)(OH)(OMe), --C(Et)(OH)(OMe), --C(Me)(OH)(OEt), and
--C(Et)(OH)(OEt).
Oxo (keto, -one): .dbd.O.
Thione (thioketone): .dbd.S.
Imino (imine): .dbd.NR, wherein R is an imino substituent, for
example, hydrogen, C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a C.sub.5-20 aryl group, preferably hydrogen or a
C.sub.1-7 alkyl group. Examples of ester groups include, but are
not limited to, .dbd.NH, .dbd.NMe, .dbd.NEt, and .dbd.NPh.
Formyl (carbaldehyde, carboxaldehyde): --C(.dbd.O)H.
Acyl (keto): --C(.dbd.O)R, wherein R is an acyl substituent, for
example, a C.sub.1-7 alkyl group (also referred to as C.sub.1-7
alkylacyl or C.sub.1-7 alkanoyl), a C.sub.3-20 heterocyclyl group
(also referred to as C.sub.3-20 heterocyclylacyl), or a C.sub.5-20
aryl group (also referred to as O.sub.5-20 arylacyl), preferably a
C.sub.1-7 alkyl group. Examples of acyl groups include, but are not
limited to, --C(.dbd.O)CH.sub.3 (acetyl),
--C(.dbd.O)CH.sub.2CH.sub.3 (propionyl),
--C(.dbd.O)C(CH.sub.3).sub.3 (t-butyryl), and --C(.dbd.O)Ph
(benzoyl, phenone).
Carboxy (carboxylic acid): --C(.dbd.O)OH.
Thiocarboxy (thiocarboxylic acid): --C(.dbd.S)SH.
Thiolocarboxy (thiolocarboxylic acid): --C(.dbd.O)SH.
Thionocarboxy (thionocarboxylic acid): --C(.dbd.S)OH.
Imidic acid: --C(.dbd.NH)OH.
Hydroxamic acid: --C(.dbd.NOH)OH.
Ester (carboxylate, carboxylic acid ester, oxycarbonyl):
--C(.dbd.O)OR, wherein R is an ester substituent, for example, a
C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl group, or a
C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl group. Examples
of ester groups include, but are not limited to,
--C(.dbd.O)OCH.sub.3, --C(.dbd.O)OCH.sub.2CH.sub.3,
--C(.dbd.O)OC(CH.sub.3).sub.3, and --C(.dbd.O)OPh.
Acyloxy (reverse ester): --OC(.dbd.O)R, wherein R is an acyloxy
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of acyloxy groups include, but are
not limited to, --OC(.dbd.O)CH.sub.3 (acetoxy),
--OC(.dbd.O)CH.sub.2CH.sub.3, --OC(.dbd.O)C(CH.sub.3).sub.3,
--OC(.dbd.O)Ph, and --OC(.dbd.O)CH.sub.2Ph.
Oxycarboyloxy: --OC(.dbd.O)OR, wherein R is an ester substituent,
for example, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a O.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl
group. Examples of ester groups include, but are not limited to,
--OC(.dbd.O)OCH.sub.3, --OC(.dbd.O)OCH.sub.2CH.sub.3,
--OC(.dbd.O)OC(CH.sub.3).sub.3, and --OC(.dbd.O)OPh.
Amino: --NR.sup.1R.sup.2, wherein R.sup.1 and R.sup.2 are
independently amino substituents, for example, hydrogen, a
C.sub.1-7 alkyl group (also referred to as C.sub.1-7 alkylamino or
di-C.sub.1-7 alkylamino), a C.sub.3-20 heterocyclyl group, or a
C.sub.5-20 aryl group, preferably H or a C.sub.1-7 alkyl group, or,
in the case of a "cyclic" amino group, R.sup.1 and R.sup.2, taken
together with the nitrogen atom to which they are attached, form a
heterocyclic ring having from 4 to 8 ring atoms. Amino groups may
be primary (--NH.sub.2), secondary (--NHR.sup.1), or tertiary
(--NHR.sup.1R.sup.2), and in cationic form, may be quaternary
(--.sup.+NR.sup.1R.sup.2R.sup.3). Examples of amino groups include,
but are not limited to, --NH.sub.2, --NHCH.sub.3,
--NHC(CH.sub.3).sub.2, --N(CH.sub.3).sub.2,
--N(CH.sub.2CH.sub.3).sub.2, and --NHPh. Examples of cyclic amino
groups include, but are not limited to, aziridino, azetidino,
pyrrolidino, piperidino, piperazino, morpholino, and
thiomorpholino.
Amido (carbamoyl, carbamyl, aminocarbonyl, carboxamide):
--C(.dbd.O)NR.sup.1R.sup.2, wherein R.sup.1 and R.sup.2 are
independently amino substituents, as defined for amino groups.
Examples of amido groups include, but are not limited to,
--C(.dbd.O)NH.sub.2, --C(.dbd.O)NHCH.sub.3,
--C(.dbd.O)N(CH.sub.3).sub.2, --C(.dbd.O)NHCH.sub.2CH.sub.3, and
--C(.dbd.O)N(CH.sub.2CH.sub.3).sub.2, as well as amido groups in
which R.sup.1 and R.sup.2, together with the nitrogen atom to which
they are attached, form a heterocyclic structure as in, for
example, piperidinocarbonyl, morpholinocarbonyl,
thiomorpholinocarbonyl, and piperazinocarbonyl.
Thioamido (thiocarbamyl): --C(.dbd.S)NR.sup.1R.sup.2, wherein
R.sup.1 and R.sup.2 are independently amino substituents, as
defined for amino groups. Examples of amido groups include, but are
not limited to, --C(.dbd.S)NH.sub.2, --C(.dbd.S)NHCH.sub.3,
--C(.dbd.S)N(CH.sub.3).sub.2, and
--C(.dbd.S)NHCH.sub.2CH.sub.3.
Acylamido (acylamino): --NR.sup.1C(.dbd.O)R.sup.2, wherein R.sup.1
is an amide substituent, for example, hydrogen, a C.sub.1-7 alkyl
group, a C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably hydrogen or a C.sub.1-7 alkyl group, and R.sup.2 is an
acyl substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20aryl group,
preferably hydrogen or a C.sub.1-7 alkyl group. Examples of
acylamide groups include, but are not limited to,
--NHC(.dbd.O)CH.sub.3, --NHC(.dbd.O)CH.sub.2CH.sub.3, and
--NHC(.dbd.O)Ph. R.sup.1 and R.sup.2 may together form a cyclic
structure, as in, for example, succinimidyl, maleimidyl, and
phthalimidyl:
##STR00010##
Aminocarbonyloxy: --OC(.dbd.O)NR.sup.1R.sup.2, wherein R.sup.1 and
R.sup.2 are independently amino substituents, as defined for amino
groups. Examples of aminocarbonyloxy groups include, but are not
limited to, --OC(.dbd.O)NH.sub.2, --OC(.dbd.O)NHMe,
--OC(.dbd.O)NMe.sub.2, and --OC(.dbd.O)NEt.sub.2.
Ureido: --N(R.sup.1)CONR.sup.2R.sup.3 wherein R.sup.2 and R.sup.3
are independently amino substituents, as defined for amino groups,
and R.sup.1 is a ureido substituent, for example, hydrogen, a
C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl group, or a
C.sub.5-20 aryl group, preferably hydrogen or a C.sub.1-7 alkyl
group. Examples of ureido groups include, but are not limited to,
--NHCONH.sub.2, --NHCONHMe, --NHCONHEt, --NHCONMe.sub.2,
--NHCONEt.sub.2, --NMeCONH.sub.2, --NMeCONHMe, --NMeCONHEt,
--NMeCONMe.sub.2, and --NMeCONEt.sub.2.
Guanidino: --NH--C(.dbd.NH)NH.sub.2.
Tetrazolyl: a five membered aromatic ring having four nitrogen
atoms and one carbon atom,
##STR00011##
Imino: .dbd.NR, wherein R is an imino substituent, for example, for
example, hydrogen, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably H or a
C.sub.1-7alkyl group. Examples of imino groups include, but are not
limited to, .dbd.NH, .dbd.NMe, and .dbd.NEt.
Amidine (amidino): --C(.dbd.NR)NR.sub.2, wherein each R is an
amidine substituent, for example, hydrogen, a C.sub.1-7 alkyl
group, a C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably H or a C.sub.1-7 alkyl group. Examples of amidine groups
include, but are not limited to, --C(.dbd.NH)NH.sub.2,
--C(.dbd.NH)NMe.sub.2, and --C(.dbd.NMe)NMe.sub.2.
Nitro: --NO.sub.2.
Nitroso: --NO.
Azido: --N.sub.3.
Cyano (nitrile, carbonitrile): --CN.
Isocyano: --NC.
Cyanato: --OCN.
Isocyanato: --NCO.
Thiocyano (thiocyanato): --SCN.
Isothiocyano (isothiocyanato): --NCS.
Sulfhydryl (thiol, mercapto): --SH.
Thioether (sulfide): --SR, wherein R is a thioether substituent,
for example, a C.sub.1-7 alkyl group (also referred to as a
C.sub.1-7alkylthio group), a O.sub.3-20 heterocyclyl group, or a
C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl group. Examples
of C.sub.1-7 alkylthio groups include, but are not limited to,
--SCH.sub.3 and --SCH.sub.2CH.sub.3.
Disulfide: --SS--R, wherein R is a disulfide substituent, for
example, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl group,
or a C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl group
(also referred to herein as C.sub.1-7 alkyl disulfide). Examples of
C.sub.1-7 alkyl disulfide groups include, but are not limited to,
--SSCH.sub.3 and --SSCH.sub.2CH.sub.3.
Sulfine (sulfinyl, sulfoxide): --S(.dbd.O)R, wherein R is a sulfine
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of sulfine groups include, but are
not limited to, --S(.dbd.O)CH.sub.3 and
--S(.dbd.O)CH.sub.2CH.sub.3.
Sulfone (sulfonyl): --S(.dbd.O).sub.2R, wherein R is a sulfone
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group, including, for example, a fluorinated or
perfluorinated C.sub.1-7 alkyl group. Examples of sulfone groups
include, but are not limited to, --S(.dbd.O).sub.2CH.sub.3
(methanesulfonyl, mesyl), --S(.dbd.O).sub.2CF.sub.3 (triflyl),
--S(.dbd.O).sub.2CH.sub.2CH.sub.3 (esyl),
--S(.dbd.O).sub.2C.sub.4F.sub.9 (nonaflyl),
--S(.dbd.O).sub.2CH.sub.2CF.sub.3 (tresyl),
--S(.dbd.O).sub.2CH.sub.2CH.sub.2NH.sub.2 (tauryl),
--S(.dbd.O).sub.2Ph (phenylsulfonyl, besyl), 4-methylphenylsulfonyl
(tosyl), 4-chlorophenylsulfonyl (closyl), 4-bromophenylsulfonyl
(brosyl), 4-nitrophenyl (nosyl), 2-naphthalenesulfonate (napsyl),
and 5-dimethylamino-naphthalen-1-ylsulfonate (dansyl).
Sulfinic acid (sulfino): --S(.dbd.O)OH, --SO.sub.2H.
Sulfonic acid (sulfo): --S(.dbd.O).sub.2OH, --SO.sub.3H.
Sulfinate (sulfinic acid ester): --S(.dbd.O)OR; wherein R is a
sulfinate substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfinate groups
include, but are not limited to, --S(.dbd.O)OCH.sub.3
(methoxysulfinyl; methyl sulfinate) and
--S(.dbd.O)OCH.sub.2CH.sub.3 (ethoxysulfinyl; ethyl sulfinate).
Sulfonate (sulfonic acid ester): --S(.dbd.O).sub.2OR, wherein R is
a sulfonate substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a O.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfonate groups
include, but are not limited to, --S(.dbd.O).sub.2OCH.sub.3
(methoxysulfonyl; methyl sulfonate) and
--S(.dbd.O).sub.2OCH.sub.2CH.sub.3 (ethoxysulfonyl; ethyl
sulfonate).
Sulfinyloxy: --OS(.dbd.O)R, wherein R is a sulfinyloxy substituent,
for example, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a C.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl
group. Examples of sulfinyloxy groups include, but are not limited
to, --OS(.dbd.O)CH.sub.3 and --OS(.dbd.O)CH.sub.2CH.sub.3.
Sulfonyloxy: --OS(.dbd.O).sub.2R, wherein R is a sulfonyloxy
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of sulfonyloxy groups include, but
are not limited to, --OS(.dbd.O).sub.2CH.sub.3 (mesylate) and
--OS(.dbd.O).sub.2CH.sub.2CH.sub.3 (esylate).
Sulfate: --OS(.dbd.O).sub.2OR; wherein R is a sulfate substituent,
for example, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a O.sub.5-20 aryl group, preferably a C.sub.1-7 alkyl
group. Examples of sulfate groups include, but are not limited to,
--OS(.dbd.O).sub.2OCH.sub.3 and
--SO(.dbd.O).sub.2OCH.sub.2CH.sub.3.
Sulfamyl (sulfamoyl; sulfinic acid amide; sulfinamide):
--S(.dbd.O)NR.sup.1R.sup.2, wherein R.sup.1 and R.sup.2 are
independently amino substituents, as defined for amino groups.
Examples of sulfamyl groups include, but are not limited to,
--S(.dbd.O)NH.sub.2, --S(.dbd.O)NH(CH.sub.3),
--S(.dbd.O)N(CH.sub.3).sub.2, --S(.dbd.O)NH(CH.sub.2CH.sub.3),
--S(.dbd.O)N(CH.sub.2CH.sub.3).sub.2, and --S(.dbd.O)NHPh.
Sulfonamido (sulfinamoyl; sulfonic acid amide; sulfonamide):
--S(.dbd.O).sub.2NR.sup.1R.sup.2, wherein R.sup.1 and R.sup.2 are
independently amino substituents, as defined for amino groups.
Examples of sulfonamido groups include, but are not limited to,
--S(.dbd.O).sub.2NH.sub.2, --S(.dbd.O).sub.2NH(CH.sub.3),
--S(.dbd.O).sub.2N(CH.sub.3).sub.2,
--S(.dbd.O).sub.2NH(CH.sub.2CH.sub.3),
--S(.dbd.O).sub.2N(CH.sub.2CH.sub.3).sub.2, and
--S(.dbd.O).sub.2NHPh.
Sulfamino: --NR.sup.1S(.dbd.O).sub.2OH, wherein R.sup.1 is an amino
substituent, as defined for amino groups. Examples of sulfamino
groups include, but are not limited to, --NHS(.dbd.O).sub.2OH and
--N(CH.sub.3)S(.dbd.O).sub.2OH.
Sulfonamino: --NR.sup.1S(.dbd.O).sub.2R, wherein R.sup.1 is an
amino substituent, as defined for amino groups, and R is a
sulfonamino substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group. Examples of sulfonamino groups
include, but are not limited to, --NHS(.dbd.O).sub.2CH.sub.3 and
--N(CH.sub.3)S(.dbd.O).sub.2C.sub.6H.sub.5.
Sulfinamino: --NR.sup.1S(.dbd.O)R, wherein R.sup.1 is an amino
substituent, as defined for amino groups, and R is a sulfinamino
substituent, for example, a C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably a
C.sub.1-7 alkyl group. Examples of sulfinamino groups include, but
are not limited to, --NHS(.dbd.O)CH.sub.3 and
--N(CH.sub.3)S(.dbd.O)C.sub.6H.sub.5.
Phosphino (phosphine): --PR.sub.2, wherein R is a phosphino
substituent, for example, --H, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably --H, a C.sub.1-7 alkyl group, or a C.sub.5-20 aryl
group. Examples of phosphino groups include, but are not limited
to, --PH.sub.2, --P(CH.sub.3).sub.2, --P(CH.sub.2CH.sub.3).sub.2,
--P(t-Bu).sub.2, and --P(Ph).sub.2.
Phosphor --P(.dbd.O).sub.2.
Phosphinyl (phosphine oxide): --P(.dbd.O)R.sub.2, wherein R is a
phosphinyl substituent, for example, a C.sub.1-7 alkyl group, a
C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably a C.sub.1-7 alkyl group or a C.sub.5-20 aryl group.
Examples of phosphinyl groups include, but are not limited to,
--P(.dbd.O)(CH.sub.3).sub.2, --P(.dbd.O)(CH.sub.2CH.sub.3).sub.2,
--P(.dbd.O)(t-Bu).sub.2, and --P(.dbd.O)(Ph).sub.2.
Phosphonic acid (phosphono): --P(.dbd.O)(OH).sub.2.
Phosphonate (phosphono ester): --P(.dbd.O)(OR).sub.2, where R is a
phosphonate substituent, for example, --H, a C.sub.1-7 alkyl group,
a C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably --H, a C.sub.1-7 alkyl group, or a C.sub.5-20 aryl
group. Examples of phosphonate groups include, but are not limited
to, --P(.dbd.O)(OCH.sub.3).sub.2,
--P(.dbd.O)(OCH.sub.2CH.sub.3).sub.2, --P(.dbd.O)(O-t-Bu).sub.2,
and --P(.dbd.O)(OPh).sub.2.
Phosphoric acid (phosphonooxy): --OP(.dbd.O)(OH).sub.2.
Phosphate (phosphonooxy ester): --OP(.dbd.O)(OR).sub.2, where R is
a phosphate substituent, for example, --H, a C.sub.1-7 alkyl group,
a C.sub.3-20 heterocyclyl group, or a C.sub.5-20 aryl group,
preferably --H, a C.sub.1-7 alkyl group, or a C.sub.5-20 aryl
group. Examples of phosphate groups include, but are not limited
to, --OP(.dbd.O)(OCH.sub.3).sub.2,
--OP(.dbd.O)(OCH.sub.2CH.sub.3).sub.2, --OP(.dbd.O)(O-t-Bu).sub.2,
and --OP(.dbd.O)(OPh).sub.2.
Phosphorous acid: --OP(OH).sub.2.
Phosphite: --OP(OR).sub.2, where R is a phosphite substituent, for
example, --H, a C.sub.1-7 alkyl group, a C.sub.3-20 heterocyclyl
group, or a C.sub.5-20 aryl group, preferably --H, a C.sub.1-7
alkyl group, or a C.sub.5-20 aryl group. Examples of phosphite
groups include, but are not limited to, --OP(OCH.sub.3).sub.2,
--OP(OCH.sub.2CH.sub.3).sub.2, --OP(O-t-Bu).sub.2, and
--OP(OPh).sub.2.
Phosphoramidite: --OP(OR.sup.1)--NR.sup.2.sub.2, where R.sup.1 and
R.sup.2 are phosphoramidite substituents, for example, --H, a
(optionally substituted) C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably --H, a
C.sub.1-7 alkyl group, or a C.sub.5-20 aryl group. Examples of
phosphoramidite groups include, but are not limited to,
--OP(OCH.sub.2CH.sub.3)--N(CH.sub.3).sub.2,
--OP(OCH.sub.2CH.sub.3)--N(i-Pr).sub.2, and
--OP(OCH.sub.2CH.sub.2CN)--N(i-Pr).sub.2.
Phosphoramidate: --OP(.dbd.O)(OR.sup.1)--NR.sup.2.sub.2, where
R.sup.1 and R.sup.2 are phosphoramidate substituents, for example,
--H, a (optionally substituted) C.sub.1-7 alkyl group, a C.sub.3-20
heterocyclyl group, or a C.sub.5-20 aryl group, preferably --H, a
C.sub.1-7 alkyl group, or a C.sub.5-20 aryl group. Examples of
phosphoramidate groups include, but are not limited to,
--OP(.dbd.O)(OCH.sub.2CH.sub.3)--N(CH.sub.3).sub.2,
--OP(.dbd.O)(OCH.sub.2CH.sub.3)--N(i-Pr).sub.2, and
--OP(.dbd.O)(OCH.sub.2CH.sub.2CN)--N(i-Pr).sub.2.
Alkylene
C.sub.3-12 alkylene: The term "C.sub.3-12 alkylene", as used
herein, pertains to a bidentate moiety obtained by removing two
hydrogen atoms, either both from the same carbon atom, or one from
each of two different carbon atoms, of a hydrocarbon compound
having from 3 to 12 carbon atoms (unless otherwise specified),
which may be aliphatic or alicyclic, and which may be saturated,
partially unsaturated, or fully unsaturated. Thus, the term
"alkylene" includes the sub-classes alkenylene, alkynylene,
cycloalkylene, etc., discussed below.
Examples of linear saturated C.sub.3-12 alkylene groups include,
but are not limited to, --(CH.sub.2).sub.n-- where n is an integer
from 3 to 12, for example, --CH.sub.2CH.sub.2CH.sub.2--
(propylene), --CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- (butylene),
--CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2-- (pentylene) and
--CH.sub.2CH.sub.2CH.sub.2CH--.sub.2CH.sub.2CH.sub.2CH.sub.2--
(heptylene).
Examples of branched saturated C.sub.3-12 alkylene groups include,
but are not limited to, --CH(CH.sub.3)CH.sub.2--,
--CH(CH.sub.3)CH.sub.2CH.sub.2--,
--CH(CH.sub.3)CH.sub.2CH.sub.2CH.sub.2--,
--CH.sub.2CH(CH.sub.3)CH.sub.2--,
--CH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2--, --CH(CH.sub.2CH.sub.3)--,
--CH(CH.sub.2CH.sub.3)CH.sub.2--, and
--CH.sub.2CH(CH.sub.2CH.sub.3)CH.sub.2--.
Examples of linear partially unsaturated C.sub.3-12 alkylene groups
(C.sub.3-12 alkenylene, and alkynylene groups) include, but are not
limited to, --CH.dbd.CH--CH.sub.2--, --CH.sub.2--CH.dbd.CH.sub.2--,
--CH.dbd.CH--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.dbd.CH--, --CH.dbd.CH--CH.dbd.CH--CH.sub.2--,
--CH.dbd.CH--CH.dbd.CH--CH.sub.2--CH.sub.2--,
--CH.dbd.CH--CH.sub.2--CH.dbd.CH--,
--CH.dbd.CH--CH.sub.2--CH.sub.2--CH.dbd.CH--, and
--CH.sub.2--C.ident.C--CH.sub.2--.
Examples of branched partially unsaturated C.sub.3-12 alkylene
groups (C.sub.3-12 alkenylene and alkynylene groups) include, but
are not limited to, --C(CH.sub.3).dbd.CH--,
--C(CH.sub.3).dbd.CH--CH.sub.2--, --CH.dbd.CH--CH(CH.sub.3)-- and
--C.ident.C--CH(CH.sub.3)--.
Examples of alicyclic saturated C.sub.3-12 alkylene groups
(C.sub.3-12 cycloalkylenes) include, but are not limited to,
cyclopentylene (e.g. cyclopent-1,3-ylene), and cyclohexylene (e.g.
cyclohex-1,4-ylene).
Examples of alicyclic partially unsaturated C.sub.3-12 alkylene
groups (C.sub.3-12 cycloalkylenes) include, but are not limited to,
cyclopentenylene (e.g. 4-cyclopenten-1,3-ylene), cyclohexenylene
(e.g. 2-cyclohexen-1,4-ylene; 3-cyclohexen-1,2-ylene;
2,5-cyclohexadien-1,4-ylene).
Oxygen protecting group: the term "oxygen protecting group" refers
to a moiety which masks a hydroxy group, and these are well known
in the art. A large number of suitable groups are described on
pages 23 to 200 of Greene, T. W. and Wuts, G. M., Protective Groups
in Organic Synthesis, 3.sup.rd Edition, John Wiley & Sons,
Inc., 1999, which is incorporated herein by reference. Classes of
particular interest include silyl ethers (e.g. TMS, TBDMS),
substituted methyl ethers (e.g. THP) and esters (e.g. acetate).
Carbamate nitrogen protecting group: the term "carbamate nitrogen
protecting group" pertains to a moiety which masks the nitrogen in
the imine bond, and these are well known in the art. These groups
have the following structure:
##STR00012## wherein R'.sup.10 is R as defined above. A large
number of suitable groups are described on pages 503 to 549 of
Greene, T. W. and Wuts, G. M., Protective Groups in Organic
Synthesis, 3.sup.rd Edition, John Wiley & Sons, Inc., 1999,
which is incorporated herein by reference.
Hemi-aminal nitrogen protecting group: the term "hemi-aminal
nitrogen protecting group" pertains to a group having the following
structure:
##STR00013## wherein R'.sup.10 is R as defined above. A large
number of suitable groups are described on pages 633 to 647 as
amide protecting groups of Greene, T. W. and Wuts, G. M.,
Protective Groups in Organic Synthesis, 3.sup.rd Edition, John
Wiley & Sons, Inc., 1999, which is incorporated herein by
reference. Methods of Treatment
The compounds of the present invention may be used in a method of
therapy. Also provided is a method of treatment, comprising
administering to a subject in need of treatment a
therapeutically-effective amount of a compound of formula I. The
term "therapeutically effective amount" is an amount sufficient to
show benefit to a patient. Such benefit may be at least
amelioration of at least one symptom. The actual amount
administered, and rate and time-course of administration, will
depend on the nature and severity of what is being treated.
Prescription of treatment, e.g. decisions on dosage, is within the
responsibility of general practitioners and other medical
doctors.
A compound may be administered alone or in combination with other
treatments, either simultaneously or sequentially dependent upon
the condition to be treated. Examples of treatments and therapies
include, but are not limited to, chemotherapy (the administration
of active agents, including, e.g. drugs; surgery; and radiation
therapy.
Pharmaceutical compositions according to the present invention, and
for use in accordance with the present invention, may comprise, in
addition to the active ingredient, i.e. a compound of formula I, a
pharmaceutically acceptable excipient, carrier, buffer, stabiliser
or other materials well known to those skilled in the art. Such
materials should be non-toxic and should not interfere with the
efficacy of the active ingredient. The precise nature of the
carrier or other material will depend on the route of
administration, which may be oral, or by injection, e.g. cutaneous,
subcutaneous, or intravenous.
Pharmaceutical compositions for oral administration may be in
tablet, capsule, powder or liquid form. A tablet may comprise a
solid carrier or an adjuvant. Liquid pharmaceutical compositions
generally comprise a liquid carrier such as water, petroleum,
animal or vegetable oils, mineral oil or synthetic oil.
Physiological saline solution, dextrose or other saccharide
solution or glycols such as ethylene glycol, propylene glycol or
polyethylene glycol may be included. A capsule may comprise a solid
carrier such a gelatin.
For intravenous, cutaneous or subcutaneous injection, or injection
at the site of affliction, the active ingredient will be in the
form of a parenterally acceptable aqueous solution which is
pyrogen-free and has suitable pH, isotonicity and stability. Those
of relevant skill in the art are well able to prepare suitable
solutions using, for example, isotonic vehicles such as Sodium
Chloride Injection, Ringer's Injection, Lactated Ringer's
Injection. Preservatives, stabilisers, buffers, antioxidants and/or
other additives may be included, as required.
Includes Other Forms
Unless otherwise specified, included in the above are the well
known ionic, salt, solvate, and protected forms of these
substituents. For example, a reference to carboxylic acid (--COON)
also includes the anionic (carboxylate) form (--COO.sup.-), a salt
or solvate thereof, as well as conventional protected forms.
Similarly, a reference to an amino group includes the protonated
form (--N.sup.+HR.sup.1R.sup.2), a salt or solvate of the amino
group, for example, a hydrochloride salt, as well as conventional
protected forms of an amino group. Similarly, a reference to a
hydroxyl group also includes the anionic form (--O.sup.-), a salt
or solvate thereof, as well as conventional protected forms.
Salts
It may be convenient or desirable to prepare, purify, and/or handle
a corresponding salt of the active compound, for example, a
pharmaceutically-acceptable salt. Examples of pharmaceutically
acceptable salts are discussed in Berge, et al., J. Pharm. Sci.,
66, 1-19 (1977).
For example, if the compound is anionic, or has a functional group
which may be anionic (e.g. --COON may be --COO.sup.-), then a salt
may be formed with a suitable cation. Examples of suitable
inorganic cations include, but are not limited to, alkali metal
ions such as Na.sup.+ and K.sup.+, alkaline earth cations such as
Ca.sup.2+ and Mg.sup.2+, and other cations such as Al.sup.+3.
Examples of suitable organic cations include, but are not limited
to, ammonium ion (i.e. NH.sub.4.sup.+) and substituted ammonium
ions (e.g. NH.sub.3R.sup.+, NH.sub.2R.sub.2.sup.+, NHR.sub.3.sup.+,
NR.sub.4.sup.+). Examples of some suitable substituted ammonium
ions are those derived from: ethylamine, diethylamine,
dicyclohexylamine, triethylamine, butylamine, ethylenediamine,
ethanolamine, diethanolamine, piperazine, benzylamine,
phenylbenzylamine, choline, meglumine, and tromethamine, as well as
amino acids, such as lysine and arginine. An example of a common
quaternary ammonium ion is N(CH.sub.3).sub.4.sup.+.
If the compound is cationic, or has a functional group which may be
cationic (e.g. --NH.sub.2 may be --NH.sub.3.sup.+), then a salt may
be formed with a suitable anion. Examples of suitable inorganic
anions include, but are not limited to, those derived from the
following inorganic acids: hydrochloric, hydrobromic, hydroiodic,
sulfuric, sulfurous, nitric, nitrous, phosphoric, and
phosphorous.
Examples of suitable organic anions include, but are not limited
to, those derived from the following organic acids:
2-acetyoxybenzoic, acetic, ascorbic, aspartic, benzoic,
camphorsulfonic, cinnamic, citric, edetic, ethanedisulfonic,
ethanesulfonic, fumaric, glucheptonic, gluconic, glutamic,
glycolic, hydroxymaleic, hydroxynaphthalene carboxylic, isethionic,
lactic, lactobionic, lauric, maleic, malic, methanesulfonic, mucic,
oleic, oxalic, palmitic, pamoic, pantothenic, phenylacetic,
phenylsulfonic, propionic, pyruvic, salicylic, stearic, succinic,
sulfanilic, tartaric, toluenesulfonic, and valeric. Examples of
suitable polymeric organic anions include, but are not limited to,
those derived from the following polymeric acids: tannic acid,
carboxymethyl cellulose.
Solvates
It may be convenient or desirable to prepare, purify, and/or handle
a corresponding solvate of the active compound. The term "solvate"
is used herein in the conventional sense to refer to a complex of
solute (e.g. active compound, salt of active compound) and solvent.
If the solvent is water, the solvate may be conveniently referred
to as a hydrate, for example, a mono-hydrate, a di-hydrate, a
tri-hydrate, etc.
Carbinolamines
The invention includes compounds where a solvent adds across the
imine bond of the PBD moiety, which is illustrated below where the
solvent is water or an alcohol (R.sup.AOH, where R.sup.A is
C.sub.1-4 alkyl):
##STR00014##
These forms can be called the carbinolamine and carbinolamine ether
forms of the PBD. The balance of these equilibria depend on the
conditions in which the compounds are found, as well as the nature
of the moiety itself.
These particular compounds may be isolated in solid form, for
example, by lyophilisation.
Isomers
Certain compounds may exist in one or more particular geometric,
optical, enantiomeric, diasteriomeric, epimeric, atropic,
stereoisomeric, tautomeric, conformational, or anomeric forms,
including but not limited to, cis- and trans-forms; E- and Z-forms;
c-, t-, and r-forms; endo- and exo-forms; R-, S-, and meso-forms;
D- and L-forms; d- and I-forms; (+) and (-) forms; keto-, enol-,
and enolate-forms; syn- and anti-forms; synclinal- and
anticlinal-forms; .alpha.- and .beta.-forms; axial and equatorial
forms; boat-, chair-, twist-, envelope-, and halfchair-forms; and
combinations thereof, hereinafter collectively referred to as
"isomers" (or "isomeric forms").
Note that, except as discussed below for tautomeric forms,
specifically excluded from the term "isomers", as used herein, are
structural (or constitutional) isomers (i.e. isomers which differ
in the connections between atoms rather than merely by the position
of atoms in space). For example, a reference to a methoxy group,
--OCH.sub.3, is not to be construed as a reference to its
structural isomer, a hydroxymethyl group, --CH.sub.2OH. Similarly,
a reference to ortho-chlorophenyl is not to be construed as a
reference to its structural isomer, meta-chlorophenyl. However, a
reference to a class of structures may well include structurally
isomeric forms falling within that class (e.g. C.sub.1-7 alkyl
includes n-propyl and iso-propyl; butyl includes n-, iso-, sec-,
and tert-butyl; methoxyphenyl includes ortho-, meta-, and
para-methoxyphenyl).
The above exclusion does not pertain to tautomeric forms, for
example, keto-, enol-, and enolate-forms, as in, for example, the
following tautomeric pairs: keto/enol (illustrated below),
imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime,
thioketone/enethiol, N-nitroso/hyroxyazo, and nitro/aci-nitro.
##STR00015##
Note that specifically included in the term "isomer" are compounds
with one or more isotopic substitutions. For example, H may be in
any isotopic form, including .sup.1H, .sup.2H (D), and .sup.3H (T);
C may be in any isotopic form, including .sup.12C, .sup.13C, and
.sup.14C; O may be in any isotopic form, including .sup.16O and
.sup.18O; and the like.
Unless otherwise specified, a reference to a particular compound
includes all such isomeric forms, including (wholly or partially)
racemic and other mixtures thereof. Methods for the preparation
(e.g. asymmetric synthesis) and separation (e.g. fractional
crystallisation and chromatographic means) of such isomeric forms
are either known in the art or are readily obtained by adapting the
methods taught herein, or known methods, in a known manner.
General Synthetic Routes
The synthesis of PBD compounds is extensively discussed in the
following references, which discussions are incorporated herein by
reference:
a) WO 00/12508 (pages 14 to 30);
b) WO 2005/023814 (pages 3 to 10);
c) WO 2004/043963 (pages 28 to 29); and
d) WO 2005/085251 (pages 30 to 39).
Synthesis Route
The compounds of the present invention, where R.sup.10 and R.sup.11
form a nitrogen-carbon double bond between the nitrogen and carbon
atoms to which they are bound, can be synthesised from a compound
of Formula 2:
##STR00016## where R.sup.2, R.sup.6, R.sup.7, R.sup.9, R.sup.6',
R.sup.7', R.sup.9', R.sup.12, X, X' and R'' are as defined for
compounds of formula I, Prot.sup.N is a nitrogen protecting group
for synthesis and Prot.sup.O is a protected oxygen group for
synthesis or an oxo group, by deprotecting the imine bond by
standard methods.
The compound produced may be in its carbinolamine or carbinolamine
ether form depending on the solvents used. For example if
Prot.sup.N is Alloc and Prot.sup.O is an oxygen protecting group
for synthesis, then the deprotection is carried using palladium to
remove the N10 protecting group, followed by the elimination of the
oxygen protecting group for synthesis. If Prot.sup.N is Troc and
Prot.sup.O is an oxygen protecting group for synthesis, then the
deprotection is carried out using a Cd/Pb couple to yield the
compound of formula (I). If Prot.sup.N is SEM, or an analogous
group, and Prot.sup.O is an oxo group, then the oxo group can be
removed by reduction, which leads to a protected carbinolamine
intermediate, which can then be treated to remove the SEM
protecting group, followed by the elimination of water. The
reduction of the compound of Formula 2 can be accomplished by, for
example, lithium tetraborohydride, whilst a suitable means for
removing the SEM protecting group is treatment with silica gel.
Compounds of formula 2 can be synthesised from a compound of
formula 3a:
##STR00017## where R.sup.2, R.sup.6, R.sup.7, R.sup.9, R.sup.6',
R.sup.7', R.sup.9', X, X' and R'' are as defined for compounds of
formula 2, by coupling an organometallic derivative comprising
R.sup.12, such as an organoboron derivative. The organoboron
derivative may be a boronate or boronic acid.
Compounds of formula 2 can be synthesised from a compound of
formula 3b:
##STR00018## where R.sup.12, R.sup.6, R.sup.7, R.sup.9, R.sup.6',
R.sup.7', R.sup.9', X, X' and R'' are as defined for compounds of
formula 2, by coupling an organometallic derivative comprising
R.sup.2, such as an organoboron derivative. The organoboron
derivative may be a boronate or boronic acid.
Compounds of formulae 3a and 3b can be synthesised from a compound
of formula 4:
##STR00019## where R.sup.2, R.sup.6, R.sup.7, R.sup.9, R.sup.6',
R.sup.7', R.sup.9', X, X' and R'' are as defined for compounds of
formula 2, by coupling about a single equivalent (e.g. 0.9 or 1 to
1.1 or 1.2) of an organometallic derivative, such as an organoboron
derivative, comprising R.sup.2 or R.sup.12.
The couplings described above are usually carried out in the
presence of a palladium catalyst, for example Pd(PPh.sub.3).sub.4,
Pd(OCOCH.sub.3).sub.2, PdCl.sub.2, Pd.sub.2(dba).sub.3. The
coupling may be carried out under standard conditions, or may also
be carried out under microwave conditions.
The two coupling steps are usually carried out sequentially. They
may be carried out with or without purification between the two
steps. If no purification is carried out, then the two steps may be
carried out in the same reaction vessel. Purification is usually
required after the second coupling step. Purification of the
compound from the undesired by-products may be carried out by
column chromatography or ion-exchange separation.
The synthesis of compounds of formula 4 where Prot.sup.O is an oxo
group and Prot.sup.N is SEM are described in detail in WO 00/12508,
which is incorporated herein by reference. In particular, reference
is made to scheme 7 on page 24, where the above compound is
designated as intermediate P. This method of synthesis is also
described in WO 2004/043963.
The synthesis of compounds of formula 4 where Prot.sup.O is a
protected oxygen group for synthesis are described in WO
2005/085251, which synthesis is herein incorporated by
reference.
Compounds of formula I where R.sup.10 and R.sup.10' are H and
R.sup.11 and R.sup.11' are SON, can be synthesised from compounds
of formula I where R.sup.10 and R.sup.11 form a nitrogen-carbon
double bond between the nitrogen and carbon atoms to which they are
bound, by the addition of the appropriate bisulphite salt or
sulphinate salt, followed by an appropriate purification step.
Further methods are described in GB 2 053 894, which is herein
incorporated by reference.
Nitrogen Protecting Groups for Synthesis
Nitrogen protecting groups for synthesis are well known in the art.
In the present invention, the protecting groups of particular
interest are carbamate nitrogen protecting groups and hemi-aminal
nitrogen protecting groups.
Carbamate nitrogen protecting groups have the following
structure:
##STR00020## wherein R'.sup.10 is R as defined above. A large
number of suitable groups are described on pages 503 to 549 of
Greene, T. W. and Wuts, G. M., Protective Groups in Organic
Synthesis, 3.sup.rd Edition, John Wiley & Sons, Inc., 1999,
which is incorporated herein by reference.
Particularly preferred protecting groups include Troc, Teoc, Fmoc,
BOC, Doc, Hoc, TcBOC, 1-Adoc and 2-Adoc.
Other possible groups are nitrobenzyloxycarbonyl (e.g.
4-nitrobenzyloxycarbonyl) and
2-(phenylsulphonyl)ethoxycarbonyl.
Those protecting groups which can be removed with palladium
catalysis are not preferred, e.g. Alloc.
Hemi-aminal nitrogen protecting groups have the following
structure:
##STR00021## wherein R'.sup.10 is R as defined above. A large
number of suitable groups are described on pages 633 to 647 as
amide protecting groups of Greene, T. W. and Wuts, G. M.,
Protective Groups in Organic Synthesis, 3.sup.rd Edition, John
Wiley & Sons, Inc., 1999, which is incorporated herein by
reference. The groups disclosed herein can be applied to compounds
of the present invention. Such groups include, but are not limited
to, SEM, MOM, MTM, MEM, BOM, nitro or methoxy substituted BOM,
Cl.sub.3CCH.sub.2OCH.sub.2--. Protected Oxygen Group for
Synthesis
Protected oxygen group for synthesis are well known in the art. A
large number of suitable oxygen protecting groups are described on
pages 23 to 200 of Greene, T. W. and Wuts, G. M., Protective Groups
in Organic Synthesis, 3.sup.rd Edition, John Wiley & Sons,
Inc., 1999, which is incorporated herein by reference.
Classes of particular interest include silyl ethers, methyl ethers,
alkyl ethers, benzyl ethers, esters, acetates, benzoates,
carbonates, and sulfonates.
Preferred oxygen protecting groups include acetates, TBS and
THP.
Further Preferences
The following preferences may apply to all aspects of the invention
as described above, or may relate to a single aspect. The
preferences may be combined together in any combination.
In some embodiments, R.sup.6', R.sup.7', R.sup.9', R.sup.10',
R.sup.11' and Y' are preferably the same as R.sup.6, R.sup.7,
R.sup.9, R.sup.10, R.sup.11 and Y respectively.
Dimer Link
Y and Y' are preferably O.
R'' is preferably a C.sub.3-7 alkylene group with no substituents.
More preferably R'' is a C.sub.3, C.sub.5 or C.sub.7 alkylene.
R.sup.6 to R.sup.9
R.sup.9 is preferably H.
R.sup.6 is preferably selected from H, OH, OR, SH, NH.sub.2, nitro
and halo, and is more preferably H or halo, and most preferably is
H.
R.sup.7 is preferably selected from H, OH, OR, SH, SR, NH.sub.2,
NHR, NRR', and halo, and more preferably independently selected
from H, OH and OR, where R is preferably selected from optionally
substituted C.sub.1-7 alkyl, C.sub.3-10 heterocyclyl and C.sub.5-10
aryl groups. R may be more preferably a C.sub.1-4 alkyl group,
which may or may not be substituted. A substituent of interest is a
C.sub.5-6 aryl group (e.g. phenyl). Particularly preferred
substituents at the 7-positions are OMe and OCH.sub.2Ph.
These preferences apply to R.sup.9', R.sup.6' and R.sup.7'
respectively.
R.sup.2
A in R.sup.2 may be phenyl group or a C.sub.5-7 heteroaryl group,
for example furanyl, thiophenyl and pyridyl. In some embodiments, A
is preferably phenyl. In other embodiments, A is preferably
thiophenyl, for example, thiophen-2-yl and thiophen-3-yl.
X is a group selected from the list comprising: OH, SH, CO.sub.2H,
COH, N.dbd.C.dbd.O and NHR.sup.N, wherein R.sup.N is selected from
the group comprising H and C.sub.1-4 alkyl. X may preferably be:
OH, SH, CO.sub.2H, --N.dbd.C.dbd.O or NH.sub.2, and may more
preferably be: OH, SH, or NH.sub.2, and most preferably is
NH.sub.2.
Q.sup.2-X may be on any of the available ring atoms of the
C.sub.5-7 aryl group, but is preferably on a ring atom that is not
adjacent the bond to the remainder of the compound, i.e. it is
preferably .beta. or .gamma. to the bond to the remainder of the
compound. Therefore, where the C.sub.5-7 aryl group (A) is phenyl,
the substituent (Q.sup.2-X) is preferably in the meta- or
para-positions, and more preferably is in the para-position.1
In some embodiments, Q.sup.1 is a single bond. In these
embodiments, Q.sup.2 is selected from a single bond and
--Z--(CH.sub.2).sub.n--, where Z is selected from a single bond, O,
S and NH and is from 1 to 3. In some of these embodiments, Q.sup.2
is a single bond. In other embodiments, Q.sup.2 is
--Z--(CH.sub.2).sub.n--. In these embodiments, Z may be O or S and
n may be 1 or n may be 2. In other of these embodiments, Z may be a
single bond and n may be 1.
In other embodiments, Q.sup.1 is --CH.dbd.CH--.
In some embodiments, R.sup.2 may be -A-CH.sub.2--X and -A-X. In
these embodiments, X may be OH, SH, CO.sub.2H, COH and NH.sub.2. In
particularly preferred embodiments, X may be NH.sub.2.
R.sup.12
R.sup.12 may be a C.sub.5-7 aryl group. A C.sub.5-7 aryl group may
be a phenyl group or a C.sub.5-7 heteroaryl group, for example
furanyl, thiophenyl and pyridyl. In some embodiments, R.sup.12 is
preferably phenyl. In other embodiments, R.sup.12 is preferably
thiophenyl, for example, thiophen-2-yl and thiophen-3-yl.
R.sup.12 may be a C.sub.8-10 aryl, for example a quinolinyl or
isoquinolinyl group. The quinolinyl or isoquinolinyl group may be
bound to the PBD core through any available ring position. For
example, the quinolinyl may be quinolin-2-yl, quinolin-3-yl,
quinolin-4-yl, quinolin-5-yl, quinolin-6-yl, quinolin-7-yl and
quinolin-8-yl. Of these quinolin-3-yl and quinolin-6-yl may be
preferred. The isoquinolinyl may be isoquinolin-1-yl,
isoquinolin-3-yl, isoquinolin-4-yl, isoquinolin-5-yl,
isoquinolin-6-yl, isoquinolin-7-yl and isoquinolin-8-yl. Of these
isoquinolin-3-yl and isoquinolin-6-yl may be preferred.
R.sup.12 may bear any number of substituent groups. It preferably
bears from 1 to 3 substituent groups, with 1 and 2 being more
preferred, and singly substituted groups being most preferred. The
substituents may be any position.
Where R.sup.12 is C.sub.5-7 aryl group, a single substituent is
preferably on a ring atom that is not adjacent the bond to the
remainder of the compound, i.e. it is preferably .beta. or .gamma.
to the bond to the remainder of the compound. Therefore, where the
C.sub.5-7 aryl group is phenyl, the substituent is preferably in
the meta- or para-positions, and more preferably is in the
para-position.
Where R.sup.12 is a C.sub.8-10 aryl group, for example quinolinyl
or isoquinolinyl, it may bear any number of substituents at any
position of the quinoline or isoquinoline rings. In some
embodiments, it bears one, two or three substituents, and these may
be on either the proximal and distal rings or both (if more than
one substituent).
R.sup.12 Substituents
If a substituent on R.sup.12 is halo, it is preferably F or Cl,
more preferably Cl.
If a substituent on R.sup.12 is ether, it may in some embodiments
be an alkoxy group, for example, a C.sub.1-7 alkoxy group (e.g.
methoxy, ethoxy) or it may in some embodiments be a C.sub.5-7
aryloxy group (e.g. phenoxy, pyridyloxy, furanyloxy). The alkoxy
group may itself be further substituted, for example by an amino
group (e.g. dimethylamino).
If a substituent on R.sup.12 is C.sub.1-7 alkyl, it may preferably
be a C.sub.1-4 alkyl group (e.g. methyl, ethyl, propryl,
butyl).
If a substituent on R.sup.12 is C.sub.3-7 heterocyclyl, it may in
some embodiments be C.sub.6 nitrogen containing heterocyclyl group,
e.g. morpholino, thiomorpholino, piperidinyl, piperazinyl. These
groups may be bound to the rest of the PBD moiety via the nitrogen
atom. These groups may be further substituted, for example, by
C.sub.1-4 alkyl groups. If the C.sub.6 nitrogen containing
heterocyclyl group is piperazinyl, the said further substituent may
be on the second nitrogen ring atom.
If a substituent on R.sup.12 is bis-oxy-C.sub.1-3 alkylene, this is
preferably bis-oxy-methylene or bis-oxy-ethylene.
Particularly preferred substituents for R.sup.12 include methoxy,
ethoxy, fluoro, chloro, cyano, bis-oxy-methylene,
methyl-piperazinyl, morpholino and methyl-thiophenyl. Another
particularly preferred substituent for R.sup.12 is
dimethylaminopropyloxy.
R.sup.12 Groups
Particularly preferred substituted R.sup.12 groups include, but are
not limited to, 4-methoxy-phenyl, 3-methoxyphenyl, 4-ethoxy-phenyl,
3-ethoxy-phenyl, 4-fluoro-phenyl, 4-chloro-phenyl,
3,4-bisoxymethylene-phenyl, 4-methylthiophenyl, 4-cyanophenyl,
4-phenoxyphenyl, quinolin-3-yl and quinolin-6-yl, isoquinolin-3-yl
and isoquinolin-6-yl, 2-thienyl, 2-furanyl, methoxynaphthyl, and
naphthyl. Another possible substituted R.sup.12 group is
4-nitrophenyl.
M and z
It is preferred that M and M' are monovalent pharmaceutically
acceptable cations, and are more preferably Na.sup.+.
z is preferably 3.
3.sup.rd Aspect
The preferences expressed above for the first aspect may apply to
the compounds of this aspect, where appropriate.
When R.sup.13 is carbamate nitrogen protecting group, it may
preferably be Teoc, Fmoc and Troc, and may more preferably be
Troc.
When R.sup.11 is O-Prot.sup.O, wherein Prot.sup.O is an oxygen
protecting group, Prot.sup.O may preferably be TBS or THP, and may
more preferably be TBS.
When R.sup.10 is a hemi-aminal nitrogen protecting group, it may
preferably be MOM, BOM or SEM, and may more preferably be SEM.
EXAMPLES
General Experimental Methods
Optical rotations were measured on an ADP 220 polarimeter
(Bellingham Stanley Ltd.) and concentrations (c) are given in g/100
mL. Melting points were measured using a digital melting point
apparatus (Electrothermal). IR spectra were recorded on a
Perkin-Elmer Spectrum 1000 FT IR Spectrometer. .sup.1H and .sup.13C
NMR spectra were acquired at 300 K using a Bruker Avance NMR
spectrometer at 400 and 100 MHz, respectively. Chemical shifts are
reported relative to TMS (.delta.=0.0 ppm), and signals are
designated as s (singlet), d (doublet), t (triplet), dt (double
triplet), dd (doublet of doublets), ddd (double doublet of
doublets) or m (multiplet), with coupling constants given in Hertz
(Hz). Mass spectroscopy (MS) data were collected using a Waters
Micromass ZQ instrument coupled to a Waters 2695 HPLC with a Waters
2996 PDA. Waters Micromass ZQ parameters used were: Capillary (kV),
3.38; Cone (V), 35; Extractor (V), 3.0; Source temperature
(.degree. C.), 100; Desolvation Temperature (.degree. C.), 200;
Cone flow rate (L/h), 50; De-solvation flow rate (L/h), 250.
High-resolution mass spectroscopy (HRMS) data were recorded on a
Waters Micromass QTOF Global in positive W-mode using metal-coated
borosilicate glass tips to introduce the samples into the
instrument. Thin Layer Chromatography (TLC) was performed on silica
gel aluminium plates (Merck 60, F.sub.254), and flash
chromatography utilised silica gel (Merck 60, 230-400 mesh ASTM).
Except for the HOBt (NovaBiochem) and solid-supported reagents
(Argonaut), all other chemicals and solvents were purchased from
Sigma-Aldrich and were used as supplied without further
purification. Anhydrous solvents were prepared by distillation
under a dry nitrogen atmosphere in the presence of an appropriate
drying agent, and were stored over 4 .ANG. molecular sieves or
sodium wire. Petroleum ether refers to the fraction boiling at
40-60.degree. C.
Compound 1b was synthesised as described in WO 00/012508 (compound
210), which is herein incorporated by reference.
General LC/MS conditions: The HPLC (Waters Alliance 2695) was run
using a mobile phase of water (A) (formic acid 0.1%) and
acetonitrile (B) (formic acid 0.1%). Gradient: initial composition
5% B over 1.0 min then 5% B to 95% B within 3 min. The composition
was held for 0.5 min at 95% B, and then returned to 5% B in 0.3
minutes. Total gradient run time equals 5 min. Flow rate 3.0
mL/min, 400 .mu.L was split via a zero dead volume tee piece which
passes into the mass spectrometer. Wavelength detection range: 220
to 400 nm. Function type: diode array (535 scans). Column:
Phenomenex.RTM. Onyx Monolithic C18 50.times.4.60 mm
LC/MS conditions specific for compounds protected by both a Troc
and a TBDMs group: Chromatographic separation of Troc and TBDMS
protected compounds was performed on a Waters Alliance 2695 HPLC
system utilizing a Onyx Monolitic reversed-phase column (3 .mu.m
particles, 50.times.4.6 mm) from Phenomenex Corp. Mobile-phase A
consisted of 5% acetonitrile--95% water containing 0.1% formic
acid, and mobile phase B consisted of 95% acetonitrile--5% water
containing 0.1% formic acid. After 1 min at 5% B, the proportion of
B was raised to 95% B over the next 2.5 min and maintained at 95% B
for a further 1 min, before returning to 95% A in 10 s and
re-equilibration for a further 50 sec, giving a total run time of
5.0 min. The flow rate was maintained at 3.0 mL/min.
LC/MS conditions specific for compound 33: LC was run on a Waters
2767 sample Manager coupled with a Waters 2996 photodiode array
detector and a Waters ZQ single quadruple mass Spectrometer. The
column used was Luna Phenyl-Hexyl 150.times.4.60 mm, 5 .mu.m, Part
no. 00E-4257-E0 (Phenomenex). The mobile phases employed were:
Mobile phase A: 100% of HPLC grade water (0.05% triethylamine),
pH=7
Mobile phase B: 20% of HPLC grade water and 80% of HPLC grade
acetonitrile (0.05% triethylamine), pH=7
The gradients used were:
TABLE-US-00001 Time Flow Rate (min) (ml/min) % A % B Initial 1.50
90 10 1.0 1.50 90 10 16.0 1.50 64 36 30.0 1.50 5 95 31.0 1.50 90 10
32.0 1.50 90 10
Mass Spectrometry was carried out in positive ion mode and SIR
(selective ion monitor) and the ion monitored was m/z=727.2.
Synthesis of Key Intermediates
##STR00022## ##STR00023##
(a)
1,1'-[[(Propane-1,3-diyl)dioxy]bis[(5-methoxy-2-nitro-1,4-phenylene)ca-
rbonyl]]bis[(2S,4R)-methyl-4-hydroxypyrrolidine-2-carboxylate]
(2a)
Method A:
A catalytic amount of DMF (2 drops) was added to a stirred solution
of the nitro-acid 1a (1.0 g, 2.15 mmol) and oxalyl chloride (0.95
mL, 1.36 g, 10.7 mmol) in dry THF (20 mL). The reaction mixture was
allowed to stir for 16 hours at room temperature and the solvent
was removed by evaporation in vacuo. The resulting residue was
re-dissolved in dry THF (20 mL) and the acid chloride solution was
added dropwise to a stirred mixture of
(2S,4R)-methyl-4-hydroxypyrrolidine-2-carboxylate hydrochloride
(859 mg, 4.73 mmol) and TEA (6.6 mL, 4.79 g, 47.3 mmol) in THF (10
mL) at -30.degree. C. (dry ice/ethylene glycol) under a nitrogen
atmosphere. The reaction mixture was allowed to warm to room
temperature and stirred for a further 3 hours after which time TLC
(95:5 v/v CHCl.sub.3/MeOH) and LC/MS (2.45 min (ES+) m/z (relative
intensity) 721 ([M+H].sup.+, 20)) revealed formation of product.
Excess THF was removed by rotary evaporation and the resulting
residue was dissolved in DCM (50 mL). The organic layer was washed
with 1N HCl (2.times.15 mL), saturated NaHCO.sub.3 (2.times.15 mL),
H.sub.2O (20 mL), brine (30 mL) and dried (MgSO.sub.4). Filtration
and evaporation of the solvent gave the crude product as a dark
coloured oil. Purification by flash chromatography (gradient
elution: 100% CHCl.sub.3 to 96:4 v/v CHCl.sub.3/MeOH) isolated the
pure amide 2a as an orange coloured glass (840 mg, 54%).
Method B:
Oxalyl chloride (9.75 mL, 14.2 g, 111 mmol) was added to a stirred
suspension of the nitro-acid 1a (17.3 g, 37.1 mmol) and DMF (2 mL)
in anhydrous DCM (200 mL). Following initial effervescence the
reaction suspension became a solution and the mixture was allowed
to stir at room temperature for 16 hours. Conversion to the acid
chloride was confirmed by treating a sample of the reaction mixture
with MeOH and the resulting bis-methyl ester was observed by LC/MS.
The majority of solvent was removed by evaporation in vacuo, the
resulting concentrated solution was re-dissolved in a minimum
amount of dry DCM and triturated with diethyl ether. The resulting
yellow precipitate was collected by filtration, washed with cold
diethyl ether and dried for 1 hour in a vacuum oven at 40.degree.
C. The solid acid chloride was added portionwise over a period of
25 minutes to a stirred suspension of
(2S,4R)-methyl-4-hydroxypyrrolidine-2-carboxylate hydrochloride
(15.2 g, 84.0 mmol) and TEA (25.7 mL, 18.7 g, 185 mmol) in DCM (150
mL) at -40.degree. C. (dry ice/CH.sub.3CN). Immediately, the
reaction was complete as judged by LC/MS (2.47 min (ES+) m/z
(relative intensity) 721 ([M+H].sup.+, 100)). The mixture was
diluted with DCM (150 mL) and washed with 1N HCl (300 mL),
saturated NaHCO.sub.3 (300 mL), brine (300 mL), filtered (through a
phase separator) and the solvent evaporated in vacuo to give the
pure product 2a as an orange solid (21.8 g, 82%).
Analytical Data:
[.alpha.].sup.22.sub.D=-46.1.degree. (c=0.47, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) (rotamers) .delta. 7.63 (s, 2H), 6.82 (s,
2H), 4.79-4.72 (m, 2H), 4.49-4.28 (m, 6H), 3.96 (s, 6H), 3.79 (s,
6H), 3.46-3.38 (m, 2H), 3.02 (d, 2H, J=11.1 Hz), 2.48-2.30 (m, 4H),
2.29-2.04 (m, 4H); .sup.13C NMR (100 MHz, CDCl.sub.3) (rotamers)
.delta. 172.4, 166.7, 154.6, 148.4, 137.2, 127.0, 109.7, 108.2,
69.7, 65.1, 57.4, 57.0, 56.7, 52.4, 37.8, 29.0; IR (ATR,
CHCl.sub.3) 3410 (br), 3010, 2953, 1741, 1622, 1577, 1519, 1455,
1429, 1334, 1274, 1211, 1177, 1072, 1050, 1008, 871 cm.sup.-1; MS
(ES.sup.+) m/z (relative intensity) 721 ([M+H].sup.+., 47), 388
(80); HRMS [M+H].sup.+. theoretical C.sub.31H.sub.36N.sub.4O.sub.16
m/z 721.2199. found (ES.sup.+) m/z 721.2227.
(a)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis[(5-methoxy-2-nitro-1,4-phenylene)ca-
rbonyl]]bis[(2S,4R)-methyl-4-hydroxypyrrolidine-2-carboxylate]
(2b)
Preparation from 1b according to Method B gave the pure product as
an orange foam (75.5 g, 82%).
Analytical Data:
(ES.sup.+) m/z (relative intensity) 749 ([M+H].sup.+., 100).
(b)
1,1'-[[(Propane-1,3-diyl)dioxy]bis(11aS,2R)-2-(hydroxy)-7-methoxy-1,2,-
3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepin-5,11-dione]
(3a)
Method A:
A suspension of 10% Pd/C (7.5 g, 10% w/w) in DMF (40 mL) was added
to a solution of the nitro-ester 2a (75 g, 104 mmol) in DMF (360
mL). The suspension was hydrogenated in a Parr hydrogenation
apparatus over 8 hours. Progress of the reaction was monitored by
LC/MS (2.12 min (ES+) m/z (relative intensity) 597 ([M+H].sup.+.,
100), (ES-) m/z (relative intensity) 595 ([M+H].sup.+., 100) after
the hydrogen uptake had stopped. Solid Pd/C was removed by
filtration and the filtrate was concentrated by rotary evaporation
under vacuum (below 10 mbar) at 40.degree. C. to afford a dark oil
containing traces of DMF and residual charcoal. The residue was
digested in EtOH (500 mL) at 40.degree. C. on a water bath (rotary
evaporator bath) and the resulting suspension was filtered through
celite and washed with ethanol (500 mL) to give a clear filtrate.
Hydrazine hydrate (10 mL, 321 mmol) was added to the solution and
the reaction mixture was heated at reflux. After 20 minutes the
formation of a white precipitate was observed and reflux was
allowed to continue for a further 30 minutes. The mixture was
allowed to cool down to room temperature and the precipitate was
retrieved by filtration, washed with diethyl ether (2*1 volume of
precipitate) and dried in a vacuum desiccator to provide 3a (50 g,
81%).
Method B:
A solution of the nitro-ester 2a (6.80 g, 9.44 mmol) in MeOH (300
mL) was added to Raney.TM. nickel (4 large spatula ends of a
.about.50% slurry in H.sub.2O) and anti-bumping granules in a
3-neck round bottomed flask. The mixture was heated at reflux and
then treated dropwise with a solution of hydrazine hydrate (5.88
mL, 6.05 g, 188 mmol) in MeOH (50 mL) at which point vigorous
effervescence was observed. When the addition was complete
(.about.30 minutes) additional Raney.TM. nickel was added carefully
until effervescence had ceased and the initial yellow colour of the
reaction mixture was discharged. The mixture was heated at reflux
for a further 30 minutes at which point the reaction was deemed
complete by TLC (90:10 v/v CHCl.sub.3/MeOH) and LC/MS (2.12 min
(ES+) m/z (relative intensity) 597 ([M+H].sup.+., 100)). The
reaction mixture was allowed to cool to around 40.degree. C. and
then excess nickel removed by filtration through a sinter funnel
without vacuum suction. The filtrate was reduced in volume by
evaporation in vacuo at which point a colourless precipitate formed
which was collected by filtration and dried in a vacuum desiccator
to provide 3a (5.40 g, 96%).
Analytical Data:
[.alpha.].sup.27.sub.D=+404.degree. (c=0.10, DMF); .sup.1H NMR (400
MHz, DMSO-d.sub.6) .delta. 10.2 (s, 2H, NH), 7.26 (s, 2H), 6.73 (s,
2H), 5.11 (d, 2H, J=3.98 Hz, OH), 4.32-4.27 (m, 2H), 4.19-4.07 (m,
6H), 3.78 (s, 6H), 3.62 (dd, 2H, J=12.1, 3.60 Hz), 3.43 (dd, 2H,
J=12.0, 4.72 Hz), 2.67-2.57 (m, 2H), 2.26 (p, 2H, J=5.90 Hz),
1.99-1.89 (m, 2H); .sup.13C NMR (100 MHz, DMSO-d.sub.6) .delta.
169.1, 164.0, 149.9, 144.5, 129.8, 117.1, 111.3, 104.5, 54.8, 54.4,
53.1, 33.5, 27.5; IR (ATR, neat) 3438, 1680, 1654, 1610, 1605,
1516, 1490, 1434, 1379, 1263, 1234, 1216, 1177, 1156, 1115, 1089,
1038, 1018, 952, 870 cm.sup.-1; MS (ES.sup.+) m/z (relative
intensity) 619 ([M+Na].sup.+., 10), 597 ([M+H].sup.+., 52), 445
(12), 326 (11); HRMS [M+H].sup.+. theoretical
C.sub.29H.sub.32N.sub.4O.sub.10 m/z 597.2191. found (ES.sup.+) m/z
597.2205.
(b)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis(11aS,2R)-2-(hydroxy)-7-methoxy-1,2,-
3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepin-5,11-dione]
(3b)
Preparation from 2b according to Method A gave the product as a
white solid (22.1 g, 86%).
Analytical Data:
MS (ES.sup.-) m/z (relative intensity) 623.3 ([M-H].sup.-.,
100);
(c)
1,1'-[[(Propane-1,3-diyl)dioxy]bis(11aS,2R)-2-(tert-butyldimethylsilyl-
oxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c][1,4]-benzodiaze-
pin-5,11-dione] (4a)
TBSCl (317 mg, 2.1 mmol) and imidazole (342 mg, 5.03 mmol) were
added to a cloudy solution of the tetralactam 3a (250 mg, 0.42
mmol) in anhydrous DMF (6 mL). The mixture was allowed to stir
under a nitrogen atmosphere for 3 hours after which time the
reaction was deemed complete as judged by LC/MS (3.90 min (ES+) m/z
(relative intensity) 825 ([M+H].sup.+., 100)). The reaction mixture
was poured onto ice (.about.25 mL) and allowed to warm to room
temperature with stirring. The resulting white precipitate was
collected by vacuum filtration, washed with H.sub.2O, diethyl ether
and dried in the vacuum desiccator to provide pure 4a (252 mg,
73%).
Analytical Data:
[.alpha.].sup.23.sub.D=+234.degree. (c=0.41, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 8.65 (s, 2H, NH), 7.44 (s, 2H),
6.54 (s, 2H), 4.50 (p, 2H, J=5.38 Hz), 4.21-4.10 (m, 6H), 3.87 (s,
6H), 3.73-3.63 (m, 4H), 2.85-2.79 (m, 2H), 2.36-2.29 (m, 2H),
2.07-1.99 (m, 2H), 0.86 (s, 18H), 0.08 (s, 12H); .sup.13C NMR (100
MHz, CDCl.sub.3) .delta. 170.4, 165.7, 151.4, 146.6, 129.7, 118.9,
112.8, 105.3, 69.2, 65.4, 56.3, 55.7, 54.2, 35.2, 28.7, 25.7, 18.0,
-4.82 and -4.86; IR (ATR, CHCl.sub.3) 3235, 2955, 2926, 2855, 1698,
1695, 1603, 1518, 1491, 1446, 1380, 1356, 1251, 1220, 1120, 1099,
1033 cm.sup.-1; MS (ES.sup.+) m/z (relative intensity) 825
([M+H].sup.+., 62), 721 (14), 440 (38); HRMS [M+H].sup.+.
theoretical C.sub.41H.sub.60N.sub.4O.sub.10Si.sub.2 m/z 825.3921.
found (ES.sup.+) m/z 825.3948.
(c)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis(11aS,2R)-2-(tert-butyldimethylsilyl-
oxy)-7-methoxy-1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c][1,4]-benzodiaze-
pin-5,11-dione] (4b)
Preparation from 3b according to the above method gave the product
as a white solid (27.3 g, 93%).
Analytical Data:
MS (ES.sup.+) m/z (relative intensity) 853.8 ([M+H].sup.+., 100),
(ES.sup.-) m/z (relative intensity) 851.6 ([M-H].sup.-., 100.
(d)
1,1'-[[(Propane-1,3-diyl)dioxy]bis(11aS,2R)-2-(tert-butyldimethylsilyl-
oxy)-7-methoxy-10-((2-(trimethylsilyl)ethoxy)methyl)-1,2,3,10,11,11a-hexah-
ydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepin-5,11-dione] (5a)
A solution of n-BuLi (4.17 mL of a 1.6 M solution in hexane, 6.67
mmol) in anhydrous THF (10 mL) was added dropwise to a stirred
suspension of the tetralactam 4a (2.20 g, 2.67 mmol) in anhydrous
THF (30 mL) at -30.degree. C. (dry ice/ethylene glycol) under a
nitrogen atmosphere. The reaction mixture was allowed to stir at
this temperature for 1 hour (now a reddish orange colour) at which
point a solution of SEMCl (1.18 mL, 1.11 g, 6.67 mmol) in anhydrous
THF (10 mL) was added dropwise. The reaction mixture was allowed to
slowly warm to room temperature and was stirred for 16 hours under
a nitrogen atmosphere. The reaction was deemed complete as judged
by TLC (EtOAc) and LC/MS (4.77 min (ES+) m/z (relative intensity)
1085 ([M+H].sup.+., 100)). The THF was removed by evaporation in
vacuo and the resulting residue dissolved in EtOAc (60 mL), washed
with H.sub.2O (20 mL), brine (20 mL), dried (MgSO.sub.4) filtered
and evaporated in vacuo to provide the crude product. Purification
by flash chromatography (80:20 v/v Hexane/EtOAc) gave the pure
N10-SEM-protected tetralactam 5a as an oil (2.37 g, 82%).
Analytical Data:
[.alpha.].sup.23.sub.D=+163.degree. (c=0.41, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.33 (s, 2H), 7.22 (s, 2H), 5.47
(d, 2H, J=9.98 Hz), 4.68 (d, 2H, J=9.99 Hz), 4.57 (p, 2H, J=5.77
Hz), 4.29-4.19 (m, 6H), 3.89 (s, 6H), 3.79-3.51 (m, 8H), 2.87-2.81
(m, 2H), 2.41 (p, 2H, J=5.81 Hz), 2.03-1.90 (m, 2H), 1.02-0.81 (m,
22H), 0.09 (s, 12H), 0.01 (s, 18H); .sup.13C NMR (100 MHz,
CDCl.sub.3) .delta. 170.0, 165.7, 151.2, 147.5, 133.8, 121.8,
111.6, 106.9, 78.1, 69.6, 67.1, 65.5, 56.6, 56.3, 53.7, 35.6, 30.0,
25.8, 18.4, 18.1, -1.24, -4.73; IR (ATR, CHCl.sub.3) 2951, 1685,
1640, 1606, 1517, 1462, 1433, 1360, 1247, 1127, 1065 cm.sup.-1; MS
(ES.sup.+) m/z (relative intensity) 1113 ([M+Na].sup.+., 48), 1085
([M+H].sup.+., 100), 1009 (5), 813 (6); HRMS [M+H].sup.+.
theoretical C.sub.53H.sub.88N.sub.4O.sub.12Si.sub.4 m/z 1085.5548.
found (ES.sup.+) m/z 1085.5542.
(d)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis(11aS,2R)-2-(tert-butyldimethylsilyl-
oxy)-7-methoxy-10-((2-(trimethylsilyl)ethoxy)methyl)-1,2,3,10,11,11a-hexah-
ydro-5H-pyrrolo[2,1-c][1,4]-benzodiazepin-5,11-dione] (5b)
Preparation from 4b according to the above method gave the product
as a pale orange foam (46.9 g, 100%), used without further
purification.
Analytical Data:
MS (ES.sup.+) m/z (relative intensity) 1114 ([M+H].sup.+., 90),
(ES) m/z (relative intensity) 1158 ([M+2Na].sup.-., 100).
(e)
1,1'-[[(Propane-1,3-diyl)dioxy]bis(11aS,2R)-2-hydroxy-7-methoxy-10-((2-
-(trimethylsilyl)ethoxy)methyl)-1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c-
][1,4]-benzodiazepin-5,11-dione] (6a)
A solution of TBAF (5.24 mL of a 1.0 M solution in THF, 5.24 mmol)
was added to a stirred solution of the bis-silyl ether 5a (2.58 g,
2.38 mmol) in THF (40 mL) at room temperature. After stirring for
3.5 hours, analysis of the reaction mixture by TLC (95:5 v/v
CHCl.sub.3/MeOH) revealed completion of reaction. The reaction
mixture was poured into a solution of saturated NH.sub.4Cl (100 mL)
and extracted with EtOAc (3.times.30 mL). The combined organic
layers were washed with brine (60 mL), dried (MgSO.sub.4), filtered
and evaporated in vacuo to provide the crude product. Purification
by flash chromatography (gradient elution: 100% CHCl.sub.3 to 96:4
v/v CHCl.sub.3/MeOH) gave the pure tetralactam 6a as a white foam
(1.78 g, 87%).
Analytical Data:
[.alpha.].sup.23.sub.D=+202.degree. (c=0.34, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.28 (s, 2H), 7.20 (s, 2H), 5.44
(d, 2H, J=10.0 Hz), 4.72 (d, 2H, J=10.0 Hz), 4.61-4.58 (m, 2H),
4.25 (t, 4H, J=5.83 Hz), 4.20-4.16 (m, 2H), 3.91-3.85 (m, 8H),
3.77-3.54 (m, 6H), 3.01 (br s, 2H, OH), 2.96-2.90 (m, 2H), 2.38 (p,
2H, J=5.77 Hz), 2.11-2.05 (m, 2H), 1.00-0.91 (m, 4H), 0.00 (s,
18H); .sup.13C NMR (100 MHz, CDCl.sub.3) .delta. 169.5, 165.9,
151.3, 147.4, 133.7, 121.5, 111.6, 106.9, 79.4, 69.3, 67.2, 65.2,
56.5, 56.2, 54.1, 35.2, 29.1, 18.4, -1.23; IR (ATR, CHCl.sub.3)
2956, 1684, 1625, 1604, 1518, 1464, 1434, 1361, 1238, 1058, 1021
cm.sup.-1; MS (ES.sup.+) m/z (relative intensity) 885
([M+29].sup.+., 70), 857 ([M+H].sup.+., 100), 711 (8), 448 (17);
HRMS [M+H].sup.+. theoretical
C.sub.41H.sub.60N.sub.4O.sub.12Si.sub.2 m/z 857.3819. found
(ES.sup.+) m/z 857.3826.
(e)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis(11aS,2R)-2-hydroxy-7-methoxy-10-((2-
-(trimethylsilyl)ethoxy)methyl)-1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,1-c-
][1,4]-benzodiazepin-5,11-dione] (6b)
Preparation from 5b according to the above method gave the product
as a white foam (15.02 g).
Analytical Data:
MS (ES.sup.+) m/z (relative intensity) 886 ([M+H].sup.+., 10),
739.6 (100), (ES.sup.-) m/z (relative intensity) 884 ([M-H].sup.-.,
40).
(f)
1,1'-[[(Propane-1,3-diyl)dioxy]bis[(11aS)-11-sulpho-7-methoxy-2-oxo-10-
-((2-(trimethylsilyl)ethoxy)methyl)1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,-
1-c][1,4]benzodiazepin-5,11-dione]] (7a)
Method A:
A 0.37 M sodium hypochlorite solution (142.5 mL, 52.71 mmol, 2.4
eq) was added dropwise to a vigorously stirred mixture of the diol
6a (18.8 g, 21.96 mmol, 1 eq), TEMPO (0.069 g, 0.44 mmol, 0.02 eq)
and 0.5 M potassium bromide solution (8.9 mL, 4.4 mmol, 0.2 eq) in
DCM (115 mL) at 0.degree. C. The temperature was maintained between
0.degree. C. and 5.degree. C. by adjusting the rate of addition.
The resultant yellow emulsion was stirred at 0.degree. C. to
5.degree. C. for 1 hour. TLC (EtOAc) and LC/MS [3.53 min. (ES+) m/z
(relative intensity) 875 ([M+Na].sup.+., 50), (ES-) m/z (relative
intensity) 852 ([M-H].sup.-., 100)] indicated that reaction was
complete.
The reaction mixture was filtered, the organic layer separated and
the aqueous layer was backwashed with DCM (.times.2). The combined
organic portions were washed with brine (.times.1), dried
(MgSO.sub.4) and evaporated to give a yellow foam. Purification by
flash column chromatography (gradient elution 35/65 v/v
n-hexane/EtOAC, 30/70 to 25/75 v/v n-hexane/EtOAC) afforded the
bis-ketone 7a as a white foam (14.1 g, 75%).
Sodium hypochlorite solution, reagent grade, available at chlorine
10-13%, was used. This was assumed to be 10% (10 g NaClO in 100 g)
and calculated to be 1.34 M in NaClO. A stock solution was prepared
from this by diluting it to 0.37 M with water. This gave a solution
of approximately pH 14. The pH was adjusted to 9.3 to 9.4 by the
addition of solid NaHCO.sub.3. An aliquot of this stock was then
used so as to give 2.4 mol eq. for the reaction.
On addition of the bleach solution an initial increase in
temperature was observed. The rate of addition was controlled, to
maintain the temperature between 0.degree. C. to 5.degree. C. The
reaction mixture formed a thick, lemon yellow coloured,
emulsion.
The oxidation was an adaptation of the procedure described in
Thomas Fey et al, J. Org. Chem., 2001, 66, 8154-8159.
Method B:
Solid TCCA (10.6 g, 45.6 mmol) was added portionwise to a stirred
solution of the alcohol 6a (18.05 g, 21.1 mmol) and TEMPO (123 mg,
0.78 mmol) in anhydrous DCM (700 mL) at 0.degree. C. (ice/acetone).
The reaction mixture was stirred at 0.degree. C. under a nitrogen
atmosphere for 15 minutes after which time TLC (EtOAc) and LC/MS
[3.57 min (ES+) m/z (relative intensity) 875 ([M+Na].sup.+., 50)]
revealed completion of reaction. The reaction mixture was filtered
through celite and the filtrate was washed with saturated aqueous
NaHCO.sub.3 (400 mL), brine (400 mL), dried (MgSO.sub.4), filtered
and evaporated in vacuo to provide the crude product. Purification
by flash column chromatography (80:20 v/v EtOAc/Hexane) afforded
the bis-ketone 7a as a foam (11.7 g, 65%).
Method C:
A solution of anhydrous DMSO (0.72 mL, 0.84 g, 10.5 mmol) in dry
DCM (18 mL) was added dropwise over a period of 25 min to a stirred
solution of oxalyl chloride (2.63 mL of a 2.0 M solution in DCM,
5.26 mmol) under a nitrogen atmosphere at -60.degree. C. (liq
N.sub.2/CHCl.sub.3). After stirring at -55.degree. C. for 20
minutes, a slurry of the substrate 6a (1.5 g, 1.75 mmol) in dry DCM
(36 mL) was added dropwise over a period of 30 min to the reaction
mixture. After stirring for a further 50 minutes at -55.degree. C.,
a solution of TEA (3.42 mL, 2.49 g; 24.6 mmol) in dry DCM (18 mL)
was added dropwise over a period of 20 min to the reaction mixture.
The stirred reaction mixture was allowed to warm to room
temperature (-1.5 h) and then diluted with DCM (50 mL). The organic
solution was washed with 1 N HCl (2.times.25 mL), H.sub.2O (30 mL),
brine (30 mL) and dried (MgSO.sub.4). Filtration and evaporation of
the solvent in vacuo afforded the crude product which was purified
by flash column chromatography (80:20 v/v EtOAc/Hexane) to afford
bis-ketone 7a as a foam (835 mg, 56%)
Analytical Data:
[.alpha.].sup.20.sub.D=+291.degree. (c=0.26, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.32 (s, 2H), 7.25 (s, 2H), 5.50
(d, 2H, J=10.1 Hz), 4.75 (d, 2H, J=10.1 Hz), 4.60 (dd, 2H, J=9.85,
3.07 Hz), 4.31-4.18 (m, 6H), 3.89-3.84 (m, 8H), 3.78-3.62 (m, 4H),
3.55 (dd, 2H, J=19.2, 2.85 Hz), 2.76 (dd, 2H, J=19.2, 9.90 Hz),
2.42 (p, 2H, J=5.77 Hz), 0.98-0.91 (m, 4H), 0.00 (s, 18H); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 206.8, 168.8, 165.9, 151.8,
148.0, 133.9, 120.9, 111.6, 107.2, 78.2, 67.3, 65.6, 56.3, 54.9,
52.4, 37.4, 29.0, 18.4, -1.24; IR (ATR, CHCl.sub.3) 2957, 1763,
1685, 1644, 1606, 1516, 1457, 1434, 1360, 1247, 1209, 1098, 1066,
1023 cm.sup.-1; MS (ES.sup.+) m/z (relative intensity) 881
([M+29].sup.+., 38), 853 ([M+H].sup.+., 100), 707 (8), 542 (12);
HRMS [M+H].sup.+. theoretical
C.sub.41H.sub.56N.sub.4O.sub.12Si.sub.2 m/z 853.3506. found
(ES.sup.+) m/z 853.3502.
(f)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis[(11aS)-11-sulpho-7-methoxy-2-oxo-10-
-((2-(trimethylsilyl)ethoxy)methyl)1,2,3,10,11,11a-hexahydro-5H-pyrrolo[2,-
1-c][1,4]benzodiazepin-5,11-dione]] (7b)
Preparation from 6b according to Method C gave the product as a
white foam (10.5 g, 76%).
Analytical Data: MS (ES.sup.+) m/z (relative intensity) 882
([M+H].sup.+., 30), 735 (100), (ES.sup.-) m/z (relative intensity)
925 ([M+45].sup.-., 100), 880 ([M-H].sup.-., 70).
(g)
1,1'-[[(Propane-1,3-diyl)dioxy]bis(11aS)-7-methoxy-2-[[(trifluoromethy-
l)sulfonyl]oxy]-10-((2-(trimethylsilyl)ethoxy)methyl)-1,10,11,11a-tetrahyd-
ro-5H-pyrrolo[2,1-c][1,4]-benzodiazepin-5,11-dione] (8a)
Anhydrous 2,6-lutidine (5.15 mL, 4.74 g, 44.2 mmol) was injected in
one portion to a vigorously stirred solution of bis-ketone 7a (6.08
g, 7.1 mmol) in dry DCM (180 mL) at -45.degree. C. (dry
ice/acetonitrile cooling bath) under a nitrogen atmosphere.
Anhydrous triflic anhydride, taken from a freshly opened ampoule
(7.2 mL, 12.08 g, 42.8 mmol), was injected rapidly dropwise, while
maintaining the temperature at -40.degree. C. or below. The
reaction mixture was allowed to stir at -45.degree. C. for 1 hour
at which point TLC (50/50 v/v n-hexane/EtOAc) revealed the complete
consumption of starting material. The cold reaction mixture was
immediately diluted with DCM (200 mL) and, with vigorous shaking,
washed with water (1.times.100 mL), 5% citric acid solution
(1.times.200 mL) saturated NaHCO.sub.3 (200 mL), brine (100 mL) and
dried (MgSO.sub.4). Filtration and evaporation of the solvent in
vacuo afforded the crude product which was purified by flash column
chromatography (gradient elution: 90:10 v/v n-hexane/EtOAc to 70:30
v/v n-hexane/EtOAc) to afford bis-enol triflate 8a as a yellow foam
(5.5 g, 70%).
Analytical Data:
[.alpha.].sup.24.sub.D=+271.degree. (c=0.18, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.33 (s, 2H), 7.26 (s, 2H), 7.14
(t, 2H, J=1.97 Hz), 5.51 (d, 2H, J=10.1 Hz), 4.76 (d, 2H, J=10.1
Hz), 4.62 (dd, 2H, J=11.0, 3.69 Hz), 4.32-4.23 (m, 4H), 3.94-3.90
(m, 8H), 3.81-3.64 (m, 4H), 3.16 (ddd, 2H, J=16.3, 11.0, 2.36 Hz),
2.43 (p, 2H, J=5.85 Hz), 1.23-0.92 (m, 4H), 0.02 (s, 18H); .sup.13C
NMR (100 MHz, CDCl.sub.3) .delta. 167.1, 162.7, 151.9, 148.0,
138.4, 133.6, 120.2, 118.8, 111.9, 107.4, 78.6, 67.5, 65.6, 56.7,
56.3, 30.8, 29.0, 18.4, -1.25; IR (ATR, CHCl.sub.3) 2958, 1690,
1646, 1605, 1517, 1456, 1428, 1360, 1327, 1207, 1136, 1096, 1060,
1022, 938, 913 cm.sup.-1; MS (ES.sup.+) m/z (relative intensity)
1144 ([M+28].sup.+., 100), 1117 ([M+H].sup.+., 48), 1041 (40), 578
(8); HRMS [M+H].sup.+. theoretical
C.sub.43H.sub.54N.sub.4O.sub.16Si.sub.2S.sub.2F.sub.6 m/z
1117.2491. found (ES.sup.+) m/z 1117.2465.
(g)
1,1'-[[(Pentane-1,5-diyl)dioxy]bis(11aS)-7-methoxy-2-[[(trifluoromethy-
l)sulfonyl]oxy]-10-((2-(trimethylsilyl)ethoxy)methyl)-1,10,11,11a-tetrahyd-
ro-5H-pyrrolo[2,1-c][1,4]-benzodiazepin-5,11-dione] (8b)
Preparation from 7b according to the above method gave the bis-enol
triflate as a pale yellow foam (6.14 g, 82%).
Analytical Data:
(ES+) m/z (relative intensity) 1146 ([M+H].sup.+., 85).
Example 1
##STR00024##
(a)
(S)-2-(4-aminophenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(trifluoromethyl-
sulfonyl)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-tet-
rahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)propoxy)-10-((2-(trime-
thylsilyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,11a-
H)-dione (9)
Solid Pd(PPh.sub.3).sub.4 (20.18 mg, 17.46 mmol) was added to a
stirred solution of the triflate 8a (975 mg, 0.87 mmol),
4-(4,4,5,5-tetramethyl-1,3,2-dioxaboralane-2-yl)aniline (172 mg,
0.79 mmol) and Na.sub.2CO.sub.3 (138 mg, 3.98 mol) in toluene (13
mL) EtOH (6.5 mL) and H.sub.2O (6.5 mL). The dark solution was
allowed to stir under a nitrogen atmosphere for 24 hours, after
which time analysis by TLC (EtOAc) and LC/MS revealed the formation
of the desired mono-coupled product and as well as the presence of
unreacted starting material. The solvent was removed by rotary
evaporation under reduced pressure and the resulting residue
partitioned between H.sub.2O (100 mL) and EtOAc (100 mL), after
eventual separation of the layers the aqueous phase was extracted
again with EtOAc (2.times.25 mL). The combined organic layers were
washed with H.sub.2O (50 mL), brine (60 mL), dried (MgSO.sub.4),
filtered and evaporated in vacuo to provide the crude Suzuki
product. The crude Suzuki product was subjected to flash
chromatography (40% EtOAc/60% Hexane.fwdarw.70% EtOAc, 30% Hexane).
Removal of the excess eluent by rotary evaporation under reduced
pressure afforded the desired product 9 (399 mg) in 43% yield.
.sup.1H-NMR: (CDCl.sub.3, 400 MHz) .delta. 7.40 (s, 1H), 7.33 (s,
1H), 7.27 (bs, 3H), 7.24 (d, 2H, J=8.5 Hz), 7.15 (t, 1H, J=2.0 Hz),
6.66 (d, 2H, J=8.5 Hz), 5.52 (d, 2H, J=10.0 Hz), 4.77 (d, 1H,
J=10.0 Hz), 4.76 (d, 1H, J=10.0 Hz), 4.62 (dd, 1H, J=3.7, 11.0 Hz),
4.58 (dd, 1H, J=3.4, 10.6 Hz), 4.29 (t, 4H, J=5.6 Hz), 4.00-3.85
(m, 8H), 3.80-3.60 (m, 4H), 3.16 (ddd, 1H, J=2.4, 11.0, 16.3 Hz),
3.11 (ddd, 1H, J=2.2, 10.5, 16.1 Hz), 2.43 (p, 2H, J=5.9 Hz),
1.1-0.9 (m, 4H), 0.2 (s, 18H). .sup.13C-NMR: (CDCl.sub.3, 100 MHz)
.delta. 169.8, 168.3, 164.0, 162.7, 153.3, 152.6, 149.28, 149.0,
147.6, 139.6, 134.8, 134.5, 127.9 (methine), 127.5, 125.1, 123.21,
121.5, 120.5 (methine), 120.1 (methine), 116.4 (methine), 113.2
(methine), 108.7 (methine), 79.8 (methylene), 79.6 (methylene),
68.7 (methylene), 68.5 (methylene), 67.0 (methylene), 66.8
(methylene), 58.8 (methine), 58.0 (methine), 57.6 (methoxy), 32.8
(methylene), 32.0 (methylene), 30.3 (methylene), 19.7 (methylene),
0.25 (methyl).
(b)
(S)-2-(4-aminophenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(4-methoxyphenyl-
)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-tetrahydro--
1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)propoxy)-10-((2-(trimethylsily-
l)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,11aH)-dione
(10)
Solid Pd(PPh.sub.3).sub.4 (10 mg, 8.69 .mu.mol) was added to a
stirred solution of the mono-triflate 9 (230 mg, 0.22 mmol) in
toluene (3 mL), EtOH (10 mL), with 4-methoxyphenyl boronic acid (43
mg, 0.28 mmol), Na.sub.2CO.sub.3 (37 mg, 0.35 mmol), in H.sub.2O
(1.5 mL) at room temperature. The reaction mixture was allowed to
stir under a nitrogen atmosphere for 20 h, at which point the
reaction was deemed complete as judged by LC/MS and TLC (EtOAc).
The solvent was removed by rotary evaporation under reduced
pressure in vacuo and the resulting residue partitioned between
EtOAc (75 mL) and H.sub.2O (75 mL). The aqueous phase was extracted
with EtOAc (3.times.30 mL) and the combined organic layers washed
with H.sub.2O (30 mL), brine (40 mL), dried (MgSO.sub.4), filtered
and evaporated to provide the crude product. The crude product was
purified by flash chromatography (60% Hexane: 40% EtOAc.fwdarw.80%
EtOAc: 20% Hexane) to provide the pure dimer as an orange foam.
Removal of the excess eluent under reduced pressure afforded the
desired product 10 (434 mg) in 74% yield.
.sup.1H-NMR: (CDCl.sub.3, 400 MHz) .delta. 7.38 (s, 2H), 7.34 (d,
2H, J=8.8 Hz), 7.30 (bs, 1H), 7.26-7.24 (m, 3H), 7.22 (d, 2H, J=8.5
Hz), 6.86 (d, 2H, J=8.8 Hz), 6.63 (d, 2H, J=8.5 Hz), 5.50 (d, 2H,
J=10.0 Hz), 4.75 (d, 1H, J=10.0 Hz), 4.74 (d, 1H, J=10.0 Hz), 4.56
(td, 2H, J=3.3, 10.1 Hz), 4.27 (t, 2H, J=5.7 Hz), 4.00-3.85 (m,
8H), 3.80 (s, 3H), 3.77-3.60 (m, 4H), 3.20-3.00 (m, 2H), 2.42 (p,
2H, J=5.7 Hz), 0.96 (t, 4H, J=8.3 Hz), 0.00 (s, 18H).
.sup.13C-NMR: (CDCl.sub.3, 100 MHz) .delta. 169.8, 169.7, 162.9,
162.7, 160.6, 152.7, 152.6, 149.0, 147.5, 134.8, 127.8 (methine),
127.4, 126.8, 125.1, 123.1, 123.0, 121.5 (methine), 120.4
(methine), 116.4 (methine), 115.5 (methine), 113.1 (methine), 108.6
(methine), 79.6 (methylene), 68.5 (methylene), 66.9 (methylene),
58.8 (methine), 57.6 (methoxy), 56.7 (methoxy), 32.8 (methylene),
30.3 (methylene), 19.7 (methylene), 0.0 (methyl).
(c)
(S)-2-(4-aminophenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(4-methoxyphenyl-
)-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-yloxy)propoxy-
)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one (11)
Fresh LiBH.sub.4 (183 mg, 8.42 mmol) was added to a stirred
solution of the SEM-dilactam 10 (428 mg, 0.42 mmol) in THF (5 mL)
and EtOH (5 mL) at room temperature. After 10 minutes, delayed
vigorous effervescence was observed requiring the reaction vessel
to be placed in an ice bath. After removal of the ice bath the
mixture was allowed to stir at room temperature for 1 hour. LC/MS
analysis at this point revealed total consumption of starting
material with very little mono-reduced product. The reaction
mixture was poured onto ice (100 mL) and allowed to warm to room
temperature with stirring. The aqueous mixture was extracted with
DCM (3.times.30 mL) and the combined organic layers washed with
H.sub.2O (20 mL), brine (30 mL) and concentrated in vacuo. The
resulting residue was treated with DCM (5 mL), EtOH (14 mL),
H.sub.2O (7 mL) and silica gel (10 g). The viscous mixture was
allowed to stir at room temperature for 3 days. The mixture was
filtered slowly through a sinter funnel and the silica residue
washed with 90% CHCl.sub.3: 10% MeOH (.about.250 mL) until UV
activity faded completely from the eluent. The organic phase was
washed with H.sub.2O (50 mL), brine 60 mL), dried (MgSO.sub.4)
filtered and evaporated in vacuo to provide the crude material. The
crude product was purified by flash chromatography (97% CHCl.sub.3:
3% MeOH) to provide the pure C2/C2' aryl PBD dimer 11 (185 mg) 61%
yield.
.sup.1H-NMR: (CDCl.sub.3, 400 MHz) .delta. 7.88 (d, 1H, J=4.0 Hz),
7.87 (d, 1H, J=4.0 Hz), 7.52 (s, 2H); 7.39 (bs, 1H), 7.37-7.28 (m,
3H), 7.20 (d, 2H, J=8.5 Hz), 6.89 (d, 2H, J=8.8 Hz), 6.87 (s, 1H),
6.86 (s, 1H), 6.67 (d, 2H, J=8.5 Hz), 4.40-4.20 (m, 6H), 3.94 (s,
6H), 3.82 (s, 3H), 3.61-3.50 (m, 2H), 3.40-3.30 (m, 2H), 2.47-2.40
(m, 2H). .sup.13C-NMR: (CDCl.sub.3, 100 MHz) .delta. 162.5 (imine
methine), 161.3, 161.1, 159.3, 156.0, 151.1, 148.1, 146.2, 140.3,
126.2 (methine), 123.2, 122.0, 120.5 (methine), 119.4, 115.2
(methine), 114.3 (methine), 111.9 (methine), 111.2 (methine), 65.5
(methylene), 56.2 (methoxy), 55.4 (methoxy), 53.9 (methine), 35.6
(methylene), 28.9 (methylene).
Example 2
##STR00025##
(a)
(S)-2-(4-aminophenyl)-7-methoxy-8-(5-((S)-7-methoxy-2-(4-methoxyphenyl-
)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-tetrahydro--
1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)pentyloxy)-10-((2-(trimethylsi-
lyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,11
aH)-dione (12)
Solid Pd(PPh.sub.3).sub.4 (32 mg, 27.7 .mu.mol) was added to a
stirred solution of the bis-triflate 8b (1.04 g, 0.91 mmol) in
toluene (10 mL), EtOH (5 mL), with 4-methoxyphenyl boronic acid
(0.202 g, 1.32 mmol), Na.sub.2CO.sub.3 (0.169 g, 1.6 mmol), in
H.sub.2O (5 mL) at 30.degree. C. The reaction mixture was allowed
to stir under a nitrogen atmosphere for 20 hours. Additional solid
4-(4,4,5,5-tetramethyl-1,3,2-dioxaboralan-2-yl)aniline (0.203 g,
0.93 mmol) and Na.sub.2CO.sub.3 (0.056 g, 0.53 mmol) were added
followed by solid Pd(PPh.sub.3).sub.4 (10 mg, 8.6 .mu.mol). The
reaction mixture was allowed to stir under a nitrogen atmosphere
for a further 20 hours. LC/MS indicated the formation of desired
product. EtOAc (100 mL) and H.sub.2O (100 mL) were added, the
aqueous was separated and extracted with EtOAc (3.times.30 mL). The
combined organic layers were washed with H.sub.2O (100 mL), brine
(100 mL), dried (MgSO.sub.4), filtered and evaporated to provide a
dark brown oil. The oil was dissolved in DCM and loaded onto a 10 g
SCX-2 cartridge pre-equilibrated with DCM (1 vol). The cartridge
was washed with DCM (3 vol), MeOH (3 vol) and the crude product
eluted with 2M NH.sub.3 in MeOH (2 vol). Flash chromatography (50%
n-hexane: 50% EtOAc.fwdarw.20% n-hexane: 80% EtOAc) provided the
pure dimer 12 as a yellow foam (0.16 g, 34%).
Analytical Data:
[.alpha.].sup.23.sub.D=+388.degree. (c=0.22, CHCl.sub.3);
.sup.1H-NMR: (CDCl.sub.3, 400 MHz) .delta. 7.39 (s, 2H), 7.35 (d,
2H, J=12.8 Hz), 7.32 (bs, 1H), 7.26-7.23 (m, 5H), 6.89 (d, 2H,
J=8.8 Hz), 6.66 (d, 2H, J=8.5 Hz), 5.55 (d, 2H, J=10.0 Hz), 4.73
(d, 1H, J=10.0 Hz), 4.72 (d, 1H, J=10.0 Hz), 4.62 (td, 2H, J=3.2,
10.4 Hz), 4.15-4.05 (m, 4H), 4.00-3.85 (m, 8H), 3.82 (s, 3H),
3.77-3.63 (m, 4H), 3.20-3.05 (m, 2H), 2.05-1.95 (m, 4H), 1.75-1.67
(m, 2H) 1.01-0.95 (m, 4H), 0.03 (s, 18H); MS (ES.sup.+) m/z
(relative intensity) 1047 ([M+H].sup.+., 45).
(b)
(S)-2-(4-aminophenyl)-7-methoxy-8-(5-((S)-7-methoxy-2-(4-methoxyphenyl-
)-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-yloxy)pentylo-
xy)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one (13)
Fresh LiBH.sub.4 (66 mg, 3.04 mmol) was added to a stirred solution
of the SEM-dilactam 12 (428 mg, 0.42 mmol) in THF (3 mL) and EtOH
(3 mL) at 0.degree. C. (ice bath). The ice bath was removed and the
reaction mixture was allowed to reach room temperature (vigorous
effervescence). After 2 hours LC/MS analysis indicated the complete
consumption of starting material. The reaction mixture was poured
onto ice (50 mL) and allowed to warm to room temperature with
stirring. The aqueous mixture was extracted with DCM (3.times.50
mL) and the combined organic layers washed with H.sub.2O (50 mL),
brine (50 mL), dried (MgSO.sub.4) and concentrated in vacuo. The
resulting residue was treated with DCM (2 mL), EtOH (5 mL),
H.sub.2O (2.5 mL) and silica gel (3.7 g). The viscous mixture was
allowed to stir at room temperature for 3 days. The mixture was
filtered through a sinter funnel and the silica residue washed with
90% CHCl.sub.3: 10% MeOH (.about.250 mL) until UV activity faded
completely from the eluent. The organic phase was dried
(MgSO.sub.4) filtered and evaporated in vacuo to provide the crude
material. The crude product was purified by flash chromatography
(99.5% CHCl.sub.3: 0.5% MeOH to 97.5% CHCl.sub.3: 2.5% MeOH in 0.5%
increments)) to provide the pure C2/C2' aryl PBD dimer 13 (59 mg,
52%).
Analytical Data:
[.alpha.].sup.28.sub.D=+760.degree. (c=0.14, CHCl.sub.3); .sup.1H
NMR (400 MHz, CDCl.sub.3) .delta. 7.89 (d, 1H, J=4.0 Hz), 7.87 (d,
1H, J=4.0 Hz), 7.52 (s, 2H), 7.39 (bs, 1H), 7.37-7.28 (m, 3H), 7.22
(d, 2H, J=8.4 Hz), 6.91 (d, 2H, J=8.8 Hz), 6.815 (s, 1H), 6.81 (s,
1H), 6.68 (d, 2H, J=8.4 Hz), 4.45-4.35 (m, 2H), 4.2-4.0 (m, 4H),
3.94 (s, 6H), 3.85-3.7 (s, 3H), 3.65-3.50 (m, 2H), 3.45-3.3 (m,
2H), 2.05-1.9 (m, 4H), 1.75-1.65 (m, 2H); MS (ES.sup.+) (relative
intensity) 754.6 ([M+H].sup.+, 100), (ES) (relative intensity)
752.5 ([M-H].sup.-., 100).
Example 3
##STR00026##
(a)
(S)-2-(thien-2-yl)-7-methoxy-8-(3-((S)-7-methoxy-2-(trifluoromethanesu-
lfonyloxy)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-te-
trahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)propyloxy)-10-((2-(tr-
imethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,-
11aH)-dione (14)
Solid Pd(PPh.sub.3).sub.4 (41 mg, 0.036 mmol) was added to a
stirred solution of the bis-triflate 8a (1 g, 0.9 mmol) in toluene
(10 mL), EtOH (5 mL), with thien-2-yl boronic acid (149 mg, 1.16
mmol), Na.sub.2CO.sub.3 (152 mg, 1.43 mmol), in H.sub.2O (5 mL).
The reaction mixture was allowed to stir under a nitrogen
atmosphere overnight at room temperature. The solvent was removed
by evaporation in vacuo and the resulting residue partitioned
between H.sub.2O (100 mL) and EtOAc (100 mL). The aqueous layer was
extracted with EtOAc (2.times.30 mL) and the combined organic
layers washed with H.sub.2O (50 mL), brine (50 mL) dried
(MgSO.sub.4), filtered and evaporated in vacuo to provide the crude
product which was purified by flash chromatography (80 hexane: 20
EtOAc.fwdarw.50 hexane: 50 EtOAc) to provide the dimmer 14 (188 mg,
20%) yield
Analytical data: LC-MS RT 4.27 mins, 1051 (M+H); .sup.1H-NMR (400
MHZ, CDCl.sub.3) .delta. 7.36 (s, 1H), 7.31 (bs, 1H), 7.27 (bs,
1H), 7.26-7.23 (m, 2H), 7.22-7.17 (m, 1H), 7.12 (bs, 1H), 7.02-6.96
(m, 2H), 5.50 (d, J=10.0 Hz, 2H), 7.75 (d, J=10.0 Hz, 2H),
4.65-4.55 (m, 2H), 4.37-4.13 (m, 4H), 4.00-3.85 (m, 8H), 3.8-3.6
(m, 4H), 3.20-3.10 (m, 2H), 2.50-2.35 (m, 2H), 1.0-0.9 (m, 4H), 0
(s, 18H).
(b)
(S)-2-(thien-2-yl)-7-methoxy-8-(3-((S)-7-methoxy-2-(trifluoromethanesu-
lfonyloxy)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-te-
trahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)pentyloxy)-10-((2-(tr-
imethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,-
11aH)-dione (15)
Solid Pd(PPh.sub.3).sub.4 (7.66 mg, 6.63 .mu.mol) was added to a
stirred, cloudy solution of 14 (174 mg, 0.17 mmol),
Na.sub.2CO.sub.3 (28 mg, 0.22 mmol) and
4-(4,4,5,5-tetramethyl-1,3,2-dioxaboralan-2-yl)aniline (47 mg, 0.22
mmol) in toluene (2-5 mL), EtOH (1.25 mL) and H.sub.2O (125 mL) at
room temperature. The reaction mixture was allowed to stir under a
N.sub.2 atmosphere for 24 hours at which point the reaction was
deemed complete by LC/MS major peak (@ 3.97 min, FW=1016, M+Na) and
TLC (EtOAc). The solvent was removed by evaporation in vacuo and
the resulting residue partitioned between EtOAc (60 mL) and
H.sub.2O (30 mL). The layers were separated and the organic phase
was washed with H.sub.2O) (20 mL), brine (30 mL) dried (MgSO.sub.4)
filtered and evaporated in vacuo to provide the crude product 123
mg, 75% yield.
Analytical data: LC-MS RT 3.98 mins, 100% area, 994 (M+H);
.sup.1H-NMR (400 MHZ, CDCl.sub.3) .delta. 7.40 (d, J=5.3 Hz, 2H),
7.30 (t, J=1.70 Hz, 1H), 7.29-7.27 (m, 3H), 7.25 (d, J=8.5 Hz, 2H),
7.21 (dd, J=1.4, 4.73 Hz, 1H), 7.03-6.97 (m, 2H), 6.66 (d, J=8.5
Hz, 2H), 5.52 (d, J=10.0 Hz, 2H), 4.78 (d, J=10.0 Hz, 1H), 4.77 (d,
J=10.0 Hz, 1H), 4.62 (dd, J=3.4, 10.5 Hz, 1H), 4.59 (dd, J=3.40,
10.6 Hz, 1H), 4.30 (t, J=5.85 Hz, 4H), 3.85-4.03 (m, 8H), 3.84-3.64
(m, 6H), 3.18 (ddd, J=2.2, 10.5, 16.0 Hz, 1H), 3.11 (ddd, J=2.2,
10.5, 16.0 Hz, 1H), 2.44 (p, J=5.85 Hz, 2H), 0.98 (t, J=1.5 Hz,
4H), 0 (s, 18H).
(c)
(S)-2-(thien-2-yl)-7-methoxy-8-(3-((S)-7-methoxy-2-(4-aminophenyl)-5-o-
xo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-yloxy)propyloxy)-1-
H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one (16)
Fresh LiBH.sub.4 (47 mg, 2.22 mmol) was added to a stirred solution
of the SEM-dilactam 15 (110 mg, 0.11 mmol) in dry THF (3 mL) and
EtOH (3 mL) at 0.degree. C. (ice bath). The ice bath was removed
and the reaction mixture stirred under a N.sub.2 atmosphere for 1
hour. Analysis of the reaction by LC/MS analysis revealed
significant formation of the desired product (Pk @2.57 min)
(I=69.32), FW=702, M+H) and half-imine. The reaction mixture was
allowed to stir for a further 1 hour after which time no further
reaction progress was observed by LC/MS. The reaction mixture was
poured onto ice, stirred and allowed to warm to room temperature.
Following partition between DCM (50 mL) and water (50 mL), the
aqueous phase was extracted with DCM (3.times.20 mL). The combined
organic layers were washed with H.sub.2O (50 mL), brine (50 mL) and
the solvent removed by evaporation in vacuo under reduced
pressure.
The resulting residue was dissolved in DCM (5 mL), EtOH (15 mL) and
H.sub.2O (7 mL) then treated with silica gel (5 g). The reaction
was allowed to stir at room temperature for 48 h. The silica was
removed by filtration through a sinter funnel and the residue
rinsed with 90:10 CHCl.sub.3: MeOH (100 mL). H.sub.2O (50 mL) was
added to the filtrate and the layers were separated (after
shaking). The aqueous layer was extracted with CHCl.sub.3
(2.times.30 mL) and H.sub.2O (50 mL), brine (50 mL), dried
(MgSO.sub.4) filtered and evaporated in vacuo to provide the crude
product. Flash chromatography (CHCl.sub.3.fwdarw.98% CHCl.sub.3: 2%
MeOH) afforded the product (41 mg, 53%).
Anayltical data: LC-MS RT 2.55 mins, 702 (M+H)
Example 4
##STR00027##
(a)
(S)-2-(4-methoxyphenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(trifluorometh-
ylsulphonyl)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a--
tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)propyloxy)-10-((2-(-
trimethylsilyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10-
H,11aH)-dione (17)
Solid 4-methoxybenzeneboronic acid (0.388 g, 2.55 mmol) was added
to a solution of the SEM protected bis triflate (8a)(3.0 g, 2.69
mmol), sodium carbonate (426 mg, 4.02 mmol) and palladium tetrakis
triphenylphosphine (0.08 mmol) in toluene (54.8 mL), ethanol (27
mL) and water (27 mL). The reaction mixture was allowed to stir at
room temperature for 3 hours. The reaction mixture was then
partitioned between ethyl acetate and water. The organic layer was
washed with water and brine and dried over magnesium sulphate.
Excess solvent was removed by rotary evaporation under reduced
pressure and the resulting residue was subjected to flash column
chromatography (silica gel; gradient elution EtOAc/hexane
30/70.fwdarw.35/65.fwdarw.40/60.fwdarw.45/55) to remove unreacted
bis-triflate (0.6 g). Removal of excess eluent from selected
fractions afforded the 4-methoxyphenyl coupled product (1.27 g,
1.18 mmol, 41%).
LC-MS RT 4.30 mins, 1076 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.41 (s, 1H), 7.39 (d, J=8.8 Hz, 2H), 7.35 (s, 1H), 7.34
(bs, 1H), 7.29 (s, 1H), 7.16 (t, J=1.9 Hz, 1H), 6.90 (d, J=8.8 Hz,
2H), 5.53 (d, J=10.0 Hz, 2H), 4.79 (d, J=10.0 Hz, 1H), 4.78 (d,
J=10.0 Hz, 1H), 4.66-4.60 (m, 2H), 4.30 (t, J=5.7 Hz, 4H), 4.0-3.94
(m, 2H), 3.93 (s, 3H), 3.92 (s, 3H), 3.84 (s, 3H), 3.83-3.60 (m,
4H), 3.22-3.10 (m, 2H), 2.45 (t, J=5.9 Hz, 2H), 1.05-0.94 (m, 4H),
0 (s, 18H).
(b)
(S)-2-(3-aminophenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(4-methoxyphenyl-
)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-tetrahydro--
1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)propyloxy)-10-((2-(trimethylsi-
lyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,11
aH)-dione (18)
Solid 3-aminobenzeneboronic acid (0.143 g, 0.92 mmol) was added to
a solution of the mono triflate (17)(0.619 g, 0.58 mmol), sodium
carbonate (195 mg, 1.84 mmol) and palladium tetrakis
triphenylphosphine (26.6 mg, 0.023 mmol) in toluene (10 mL),
ethanol (5 mL) and water (5 mL). The reaction mixture was allowed
to stir at room temperature for overnight at 30.degree. C. The
reaction mixture was then partitioned between ethyl acetate and
water. The organic layer was washed with water and brine and dried
over magnesium sulphate. Excess solvent was removed by rotary
evaporation under reduced pressure and the resulting residue was
subjected to flash column chromatography (silica gel; gradient
elution EtOAc/hexane 70/3085/15). Removal of excess eluent from
selected fractions afforded the desired product (0.502 g, 0.49
mmol, 85%).
LC-MS RT 4.02 mins, 1019 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.38-7.35 (m, 4H), 7.33 (bs, 1H), 7.30 (bs, 1H), 7.25 (s,
2H), 7.10 (t, J=7.8 Hz, 1H), 6.88-6.80 (m, 3H), 6.72 (bs, 1H), 6.57
(dd, J=7.9, 1.8 Hz, 1H), 5.50 (d, J=10.0 Hz, 2H), 4.75 (d, 10.0 Hz,
2H), 4.58 (dd, J=10.6, 3.3 Hz, 2H), 4.27 (t, J=5.8 Hz, 4H),
3.95-3.91 (m, 2H), 3.90 (s, 6H), 3.80 (s, 3H), 3.77-3.60 (m, 6H),
3.15-3.05 (m, 2H), 2.41 (p, J=5.8 Hz, 2H), 0.95 (t, =8.25 Hz, 4H),
0 (s, 18H).
(c)
(S)-2-(3-aminophenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(4-methoxyphenyl-
)-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-yloxy)propylo-
xy)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one (19)
A solution of superhydride (0.56 mL, 0.56 mmol, 1.0 M in THF) was
added dropwise to a solution of the SEM dilactam (18)(0.271 g, 0.27
mmol) in dry THF (10 mL) at -78.degree. C. under a nitrogen
atmosphere. After 1 hr a further aliquot of superhydride solution
(0.13 ml, 0.13 mmol) was added and the reaction mixture was allowed
to stir for another 0.5 hr, at which time LC-MS indicated that
reduction was complete. The reaction mixture was diluted with water
and allowed to warm to room temperature. The reaction mixture was
partitioned between chloroform and water, the layers were separated
and the aqueous layer extracted with additional chloroform
(emulsions). Finally the combined organic phase was washed with
brine and dried over magnesium sulphate. The reduced product was
dissolved in methanol, chloroform and water and allowed to stir in
the presence of silica gel for 72 hours The crude product was
subjected to flash column chromatography (methanol/chloroform
gradient) to afford the desired imine product (150 mg, 0.21 mmol,
77%) after removal of excess eluent from selected fractions.
LC-MS RT 2.63 mins, 97% area, 726 (M+H); .sup.1H-NMR (400 MHZ,
CDCl.sub.3) .delta. 7.85 (d, J=3.9 Hz, 1H), 7.84 (d, J=3.9 Hz, 1H),
7.50 (s, 1H), 7.49 (s, 1H), 7.42 (s, 1H), 7.36 (s, 1H), 7.32 (d,
J=7.3 Hz, 2H), 7.11 (t, (d, J=7.8 Hz, 1H), 6.90-6.80 (m, 4H), 6.77
(d, J=7.9 Hz, 1H), 4.40-4.20 (m, 6H), 3.92 (s, 6H), 3.80 (s, 3H),
3.60-3.27 (m, 6H), 2.48-2.29 (m, 2H)
Example 5
##STR00028##
(a) (11S,11aS)-2,2,2-trichloroethyl
11-(tert-butyldimethylsilyloxy)-8-(5-((11
S,11aS)-11-(tert-butyldimethylsilyloxy)-7-methoxy-2-(4-methoxyphenyl)-5-o-
xo-10-((2,2,2-trichloroethoxy)carbonyl)-5,10,11,11a-tetrahydro-1H-pyrrolo[-
2,1-c][1,4]benzodiazepin-8-yloxy)pentyloxy)-7-methoxy-5-oxo-2-(trifluorome-
thylsulfonyloxy)-11,11a-dihydro-pyrrolo[2,1-c][1,4]benzodiazepine-10(5H)-c-
arboxylate 21
Solid 4-methoxybenzeneboronic acid (59 mg, 0.39 mmol) was added to
a solution of the Troc protected bis triflate (Compound 44, WO
2006/111759) (600 mg, 0.41 mmol), sodium carbonate (65 mg, 0.61
mmoml) and palladium tetrakis triphenylphosphine (0.012 mmol) in
toluene (10.8 mL), ethanol (5.4 mL) and water (5.4 mL). The
reaction mixture was allowed to stir at room temperature overnight.
The reaction mixture was then partitioned between ethylacetate and
water. The organic layer was washed with water and brine and dried
over magnesium sulphate. Excess solvent was removed by rotary
evaporation under reduced pressure and the resulting residue was
subjected to flash column chromatography (silica gel; gradient
elution EtOAc/hexane 20/80.fwdarw.30/70.fwdarw.40/60.fwdarw.60/40)
to remove unreacted bis-triflate. Removal of excess eluent from
selected fractions afforded the 4-methoxyphenyl coupled product
(261 mg, 0.18 mmol, 46%).
LC-MS RT 4.17 mins, 1427 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.38 (s, 1H), 7.33 (s, 1H), 7.31 (s, 1H), 7.30 (s, 1H),
7.25 (s, 1H), 7.20 (bs, 1H), 6.92 (d, J=8.6 Hz, 2H), 6.77 (d, J=8.7
Hz, 2H), 6.0-5.90 (m, 2H), 5.25 (d, J=12.0 Hz, 1H), 5.24 (d, J=12.0
Hz, 1H), 4.24 (d, J=12.0 Hz, 1H), 4.22 (d, J=12.0 Hz, 1H),
4.18-4.08 (m, 2H), 4.07-3.89 (m, 10H), 3.81 (s, 3H), 3.44-3.25 (m,
2H), 2.85 (d, J=16.6 Hz, 2H), 2.05-1.90 (m, 4H), 1.76-1.64 (m, 2H),
0.93 (s, 9H), 0.90 (s, 9H), 0.30 (s, 6H), 0.26 (s, 6H).
(b) (11S,11aS)-2,2,2-trichloroethyl
11-(tert-butyldimethylsilyloxy)-8-(5-((11S,11aS)-11-(tert-butyldimethylsi-
lyloxy)-2-(4-hydroxyphenyl)-7-methoxy-5-oxo-10-((2,2,2-trichloroethoxy)car-
bonyl)-5,10,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-8-yloxy)-
pentyloxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-dihydro-1H-pyrrolo[2-
,1-c][1,4]benzodiazepine-10(5H)-carboxylate 22
The Suzuki coupling procedure described in step (a) was applied to
the synthesis of Compound 21. Compound 20 (62.5 mg 0.044 mmol) was
treated with 1 equivalent of 4-hydroxybenzeneboronic acid (10 mg)
at 30.degree. C. overnight to afford the desired compound after
filtration through a pad of silica gel. (40 mg, 0.029 mmol, 66%
yield). The compound was used directly in the subsequent step
LC-MS RT 4.27 mins, 1371 (M+H)
(c)
(S)-2-(4-hydroxyphenyl)-7-methoxy-8-(5-((S)-7-methoxy-2-(4-methoxyphen-
yl)-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepindiazepine-8-ylo-
xy)pentyloxy)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11 aH)-one
23
Cadmium/lead couple (100 mg, Q Dong et al. Tetrahedron Letters vol
36, issue 32, 5681-5682, 1995) was added to a solution of 21 (40
mg, 0.029 mmol) in THF (1 mL) and ammonium acetate (1N, 1 mL) and
the reaction mixture was allowed to stir for 1 hour. The reaction
mixture was partitioned between chloroform and water, the phases
separated and the aqueous phase extracted with chloroform. The
combined organic layers were washed with brine and dried over
magnesium sulphate. Rotary evaporation under reduced pressure
yielded the crude product which was subjected to column
chromatography (silica gel, 0.fwdarw.4% MeOH/CHCl.sub.3). Removal
of excess eluent by rotary evaporation under reduced pressure
afforded the desired imine product (17 mg 0.023 mmol 79%).
LC-MS RT 2.20 mins, 755 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.89 (d, J=3.94 Hz, 1H), 7.89 (d, J=4.00 Hz, 1H), 7.53 (s,
1H), 7.52 (s, 1H), 7.38 (d, J=8.7 Hz, 2H), 7.33 (d, J=8.6 Hz, 2H),
7.28 (s, 1H), 6.90 (d, J=8.7 Hz, 2H), 6.84 (d, J=8.6 Hz, 2H), 6.82
(s, 1H), 6.81 (s, 1H), 5.68 (bs, 1H), 4.50-4.30 (m, 2H), 4.22-4.00
(m, 4H), 3.93 (s, 6H), 3.82 (s, 3H), 3.69-3.45 (m, 2H), 3.44-3.28
(m, 2H), 2.64-1.88 (m, 4H), 1.77-1.62 (m, 2H).
Example 6
##STR00029##
(a) (11S,11aS)-2,2,2-trichloroethyl
11-(tert-butyldimethylsilyloxy)-8-(5-((11 S,11a
S)-11-(tert-butyldimethylsilyloxy)-2-(4-formylphenyl)-7-methoxy-5-oxo-10--
((2,2,2-trichloroethoxy)carbonyl)-5,10,11,11a-tetrahydro-1H-pyrrolo[2,1-c]-
[1,4]benzodiazepin
diazepin-8-yloxy)pentyloxy)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-11,11a-di-
hydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-10(5H)-carboxylate
24
The Suzuki coupling procedure described in Example 5, step (a), was
applied to the synthesis of Compound 24. Compound 21 (62.5 mg,
0.044 mmol) was treated with 1 equivalent of 4-formylbenzeneboronic
acid (10.5 mg) at room temperature overnight to afford the desired
compound after filtration through a pad of silica gel (45 mg, 0.033
mmol, 75% yield). The compound was used directly in the subsequent
step.
LC-MS RT 4.42 mins, 1383 (M+H)
(b)
4-((S)-7-methoxy-8-(5-((S)-7-methoxy-2-(4-methoxyphenyl)-5-oxo-5,11a-d-
ihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-yloxy)pentyloxy)-5-oxo-5,11a-
-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-2-yl)benzaldehyde
25
Compound 24 was deprotected by the method described in Example 5,
step (c), to yield the desired compound (18 mg, 0.023 mmol,
79%).
LC-MS RT 3.18 mins, 768 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 9.98 (s, 1H), 7.91 (d, J=3.90 Hz, 1H), 7.90-7.80 (m, 3H),
7.68 (s, 1H), 7.60-7.45 (m, 4H), 7.39 (s, 1H), 7.33 (d, J=8.7 Hz,
1H), 6.90 (d, J=8.7 Hz, 2H), 6.83 (s, 1H), 6.82 (s, 1H), 4.55-4.44
(m, 1H), 4.43-4.36 (m, 1H), 4.23-4.00 (m, 4H), 3.95 (s, 3H), 3.94
(s, 3H), 3.82 (s, 3H), 3.66-3.51 (m, 2H), 3.50-3.34 (m, 2H),
2.05-1.87 (m, 4H), 1.76-164 (m, 2H).
Example 7
##STR00030##
(a) (11S,11a S)-2,2,2-trichloroethyl
2-(3-aminophenyl)-11-(tert-butyldimethylsilyloxy)-8-(5-((11S,11aS)-11-(te-
rt-butyldimethylsilyloxy)-7-methoxy-5-oxo-10-((2,2,2-trichloroethoxy)carbo-
nyl)-2-(trifluoromethylsulphonyloxy)-5,10,11,11a-tetrahydro-1H-pyrrolo[2,1-
-c][1,4]benzodiazepindiazepin-8-yloxy)pentyloxy)-7-methoxy-5-oxo-11,11a-di-
hydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-10(5H)-carboxylate
26
The Suzuki coupling procedure described in Example 5, step (a), was
applied to the synthesis of Compound 26, using
3-aminobenzeneboronic acid to afford the desired compound in 41%
yield (230 mg, 0.163 mmol)
LC-MS RT 4.28 mins, 1411 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.44 (bs, 1H), 7.29 (s, 1H), 7.25 (s, 1H), 7.20 (s, 1H),
7.16 (t, J=7.9 Hz, 1H), 6.84-6.73 (m, 3H), 6.70 (bs, 1H), 6.62 (dd,
J=7.9, 1.7 Hz, 1H), 6.66-6.58 (m, 2H), 5.25 (d, J=12.0 Hz, 1H),
5.24 (d, J=12.0 Hz, 1H), 4.24 (d, J=12.0 Hz, 1H), 4.22 (d, J=12.0
Hz, 1H), 4.17-4.07 (m, 2H), 4.08-3.89 (m, 10H), 3.43-3.28 (m, 2H),
2.85 (d, J=1.65 Hz, 2H), 2.07-1.90 (m, 4H), 1.78-1.63 (m, 2H), 0.94
(s, 9H), 0.90 (s, 9H), 0.30 (s, 6H), 0.27 (s, 6H).
(b) (11S,11aS)-2,2,2-trichloroethyl
2-(3-aminophenyl)-11-(tert-butyldimethylsilyloxy)-8-(5-((11S,11aS)-11-(te-
rt-butyldimethylsilyloxy)-2-(4-(3-(dimethylamino)propoxy)phenyl)-7-methoxy-
-5-oxo-10-((2,2,2-trichloroethoxy)carbonyl)-5,10,11,11a-tetrahydro-1H-pyrr-
olo[2,1-c][1,4]benzodiazepindiazepin-8-yloxy)pentyloxy)-7-methoxy-5-oxo-11-
,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-10(5H)-carboxylate
27
Solid 4-[3-(dimethylamino)propoxybenzeneboronic acid pinacol ester
(25 mg, 0.082 mmol) was added to a solution of 26 (73 mg, 0.052
mmol mmol), sodium carbonate (18 mg, 0.17 mmol) and palladium
tetrakis triphenylphosphine (3 mg) in toluene (1 mL), ethanol (0.5
mL) and water (0.5 mL). The reaction mixture was allowed to stir at
room temperature over night. The reaction mixture was then
partitioned between ethyl acetate and water. The organic layer was
washed with water and brine and dried over magnesium sulphate.
Excess solvent was removed by rotary evaporation under reduced
pressure and the resulting residue was eluted through a plug of
silica gel with chloroform/methanol. Removal of excess eluent from
selected fractions afforded the 4-methoxyphenyl coupled product (50
mg, 0.035 mmol, 67%).
LC-MS RT 4.12 mins, 1440 (M+H)
(c)
(S)-2-(3-aminophenyl)-8-(5-((S)-2-(4-(3-(dimethylamino)propoxy)phenyl)-
-7-methoxy-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-ylox-
y)pentyloxy)-7-methoxy-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one
28
Compound 27 was deprotected by the method described in Example 5,
step (c), to yield the desired compound. The reaction mixture was
partitioned between DCM and aqueous sodium hydrogen carbonate
(emulsion) and the crude product purified by gradient column
chromatography on silica gel (5% methanol chloroform.fwdarw.35%
methanol/chloroform) to afford the desired unsymmetrical PBD imine
(50 mg, 0.018 mmol, 58%)
LC-MS RT 2.55 mins, 826 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.92-7.82 (m, 2H), 7.52 (bs, 2H), 7.45 (bs, 1H), 7.39 (bs,
1H), 7.31 (d, J=8.6 Hz, 2H), 7.14 (t, J=7.8 Hz, 1H), 6.89 (d, J=8.6
Hz, 2H), 6.85-6.75 (m, 3H), 6.72 (bs, 1H), 6.60 (d, J=8.0 Hz, 1H),
4.46-4.33 (m, 2H), 4.21-3.98 (m, 6H), 3.94 (s, 6H), 3.63-3.50 (m,
2H), 3.43-3.29 (m, 2H), 2.64-2.48 (m, 2H), 2.34 (s, 6H), 2.10-1.89
(m, 6H), 1.57 (m, 2H).
Example 8
##STR00031##
(a) (11S,11aS)-2,2,2-trichloroethyl
2-(3-aminophenyl)-11-(tert-butyldimethylsilyloxy)-8-(5-((11S,11aS)-11-(te-
rt-butyldimethylsilyloxy)-7-methoxy-2-(4-(4-methylpiperazin-1-yl)phenyl)-5-
-oxo-10-((2,2,2-trichloroethoxy)carbonyl)-5,10,11,11a-tetrahydro-1H-pyrrol-
o[2,1-c][1,4]benzodiazepin
diazepin-8-yloxy)pentyloxy)-7-methoxy-5-oxo-11,11a-dihydro-1H-pyrrolo[2,1-
-c][1,4]benzodiazepine-10(5H)-carboxylate 29
The method of Example 7, step (b), was performed to afford the
desired product (58 mg, 0.040 mmol, 78%) after filtration through a
plug of silica gel (with 1/3 methanol/chloroform) and removal of
excess solvent by rotary evaporation under reduced pressure.
LC-MS RT 4.08 mins, 1439 (M+H)
(b)
(S)-2-(3-aminophenyl)-7-methoxy-8-(5-((S)-7-methoxy-2-(4-(4-methylpipe-
razin-1-yl)phenyl)-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepin-
e-8-yloxy)pentyloxy)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one
30
The method of Example 7, step (c) was used to deprotect compound
29. The crude product was purified by silica gel gradient
chromatography (2% methanol chloroform-35% methanol/chloroform) to
afford the desired unsymmetrical PBD imine (18 mg, 0.022 mmol,
59%)
LC-MS RT 2.52 mins, 823 (M+H); .sup.1H-NMR (400 MHZ, CDCl.sub.3)
.delta. 7.80 (d, J=3.8 Hz, 2H), 7.45 (s, 2H), 7.38 (s, 1H), 7.30
(s, 1H), 7.23 (d, J=8.6 Hz, 2H), 7.07 (t, J=7.8 Hz, 1H), 6.83 (d,
J=8.6 Hz, 2H), 6.79-6.89 (m, 3H), 6.65 (s, 1H), 6.54 (d, J=7.9 Hz,
1H), 4.40-4.24 (m, 2H), 4.15-3.93 (m, 4H), 3.87 (s, 6H), 3.56-3.42
(m, 2H), 3.37-3.23 (m, 2H), 3.22-3.08 (m, 4H), 2.61-2.41 (m, 4H),
2.29 (s, 3H), 1.98-1.80 (m, 4H), 1.67-1.54 (m, 2H).
Example 9
##STR00032##
(a)
(S)-2-(4-(aminomethyl)phenyl)-7-methoxy-8-(3-((7-methoxy-2-(4-methoxyp-
henyl)-5,11-dioxo-10-((2-(trimethylsilyl)ethoxy)methyl)-5,10,11,11a-tetrah-
ydro-1H-benzo[e]pyrrolo[1,2-a][1,4]diazepin-8-yloxy)propyloxy)-10-((2-(tri-
methylsilyl)ethoxy)methyl)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,
11aH)-dione 31
Solid 4-aminomethylbenzeneboronic acid hydrochloride (0.111 g, 0.59
mmol) was added to a solution of 17 (0.394 g, 0.37 mmol), sodium
carbonate (175 mg, 1.654 mmol) and palladium tetrakis
triphenylphosphine (28.0 mg, 0.024 mmol) in toluene (10 mL),
ethanol (5 mL) and water (5 mL). The reaction mixture was allowed
to stir overnight at 30.degree. C. The following day the reaction
mixture was heated for a further 3 hours at 70.degree. C. The
reaction mixture was then partitioned between ethyl acetate and
water. The organic layer was washed with water and brine and dried
over magnesium sulphate. Excess solvent was removed by rotary
evaporation under reduced pressure and the resulting residue was
subjected to flash column chromatography (silica gel; gradient
elution EtOAc/hexane 2/98.fwdarw.05/85). Removal of excess eluent
from selected fractions afforded the desired product (0.230 mg,
0.22 mmol, 61%).
LC-MS RT 3.63 mins, 1034 (M+2H); .sup.1H-NMR (400 MHz, DMSO
d.sub.6) .delta. 11.7 (s, 2H), 7.52 (d, J=8.2 Hz, 2H), 7.48 (d,
J=8.7 Hz, 2H), 7.40 (s, 1H), 7.50 (d, J=8.1 Hz, 2H), 7.38-7.19 (m,
5H) 6.93 (d, J=8.7 Hz, 2H), 5.40 (d, J=2.13 Hz, 1H), 5.38 (d,
J=2.12 Hz, 1H), 5.32 (d, J=10.6 Hz, 2H), 5.25 (d, J=10.6 Hz, 2H),
4.87-4.72 (m, 2H), 4.35-4.15 (m, 4H), 3.85 (s, 6H), 3.79 (s, 3H),
3.73-3.56 (m, 2H), 3.55-3.39 (m, 4H), 3.22-3.02 (m, 2H), 2.39-2.23
(m, 2H), 0.94-0.67 (m, 4H), -0.06 (s, 18H).
(b)
(S)-2-(4-(aminomethyl)phenyl)-7-methoxy-8-(3-((S)-7-methoxy-2-(4-metho-
xyphenyl)-5-oxo-5,11a-dihydro-1H-pyrrolo[2,1-c][1,4]benzodiazepine-8-yloxy-
)propyloxy)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(11aH)-one 32
Compound 31 was deprotected following the method of Example 1, step
(c). The crude product was purified by gradient column
chromatography (5/95.fwdarw.30/70 MeOH/CHCl.sub.3) to afford the
product as a mixture of imine and carbinolamine methyl ethers.
LC-MS RT 2.58 mins, 740 (M+H).
Example 10
##STR00033##
(S)-2-(4-aminophenyl)-7-methoxy-11(S)-sulpho-8-(3-((S)-7-methoxy-11(S)-sul-
pho-2-(4-methoxyphenyl)-5-oxo-5,10,11,11a-tetrahydro-1H-pyrrolo[2,1-c][1,4-
]benzodiazepine-8-yloxy)propyloxy)-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5(-
11aH)-one disodium salt 33
Sodium bisulphite (8.5 mg, 3.1 eq) was added to a stirred
suspension of bis-imine 11 (20 mg, 0.036 mmol) in isopropanol (4
mL) and water (2 mL). The reaction mixture was allowed to stir
vigorously and eventually became clear (c, 1 hour). The reaction
mixture was transferred to a funnel and filtered through a cotton
wall (and then washed with 2 mL water). The filtrate was flash
frozen (liquid and to bath) and lyophilized to afford the desired
product 33 in quantitative yield.
LC-MS RT 11.77 mins, 727.2 (M+H) (Mass of parent compound,
bisulphite adducts unstable in mass spectrometer); .sup.1H-NMR (400
MHz, CDCl.sub.3) .delta. 7.66-7.55 (m, 5H), 7.43 (s, 1H), 7.39 (d,
J=8.66 Hz, 2H), 7.06 (m, 2H), 6.93 (d, J=8.84 Hz, 2H), 6.54 (m,
2H), 5.29-5.21 (m, 2H), 4.32-4.28 (m, 2H), 4.14-4.20 (m, 4H),
3.96-3.83 (m, 2H), 3.77 (s, 3H), 3.73 (m, 6H), 3.52-3.43 (m, 2H),
3.30-3.08 (m, 2H), 2.24-2.21 (m, 2H).
Example 11
Determination of In Vitro Cytotoxicity
K562 Assay
K562 human chronic myeloid leukaemia cells were maintained in RPM1
1640 medium supplemented with 10% fetal calf serum and 2 mM
glutamine at 37.degree. C. in a humidified atmosphere containing 5%
CO.sub.2 and were incubated with a specified dose of drug for 1
hour or 96 hours at 37.degree. C. in the dark. The incubation was
terminated by centrifugation (5 min, 300 g) and the cells were
washed once with drug-free medium. Following the appropriate drug
treatment, the cells were transferred to 96-well microtiter plates
(10.sup.4 cells per well, 8 wells per sample). Plates were then
kept in the dark at 37.degree. C. in a humidified atmosphere
containing 5% CO.sub.2. The assay is based on the ability of viable
cells to reduce a yellow soluble tetrazolium salt,
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide
(MTT, Aldrich-Sigma), to an insoluble purple formazan precipitate.
Following incubation of the plates for 4 days (to allow control
cells to increase in number by approximately 10 fold), 20 .mu.L of
MTT solution (5 mg/mL in phosphate-buffered saline) was added to
each well and the plates further incubated for 5 h. The plates were
then centrifuged for 5 min at 300 g and the bulk of the medium
pipetted from the cell pellet leaving 10-20 .mu.L per well. DMSO
(200 .mu.L) was added to each well and the samples agitated to
ensure complete mixing. The optical density was then read at a
wavelength of 550 nm on a Titertek Multiscan ELISA plate reader,
and a dose-response curve was constructed. For each curve, an
IC.sub.50 value was read as the dose required to reduce the final
optical density to 50% of the control value.
Compound 13 has an IC.sub.50 of 30 .mu.M in this assay.
A2780 assay
A2780 parental cell line was grown in Dulbecco's Modified Eagles'
Media (DMEM) containing .about.10% Foetal Calf Serum (FCS) and
.about.1% 200 mM L-Glutamine solution and grown in Corning Cellbind
75 cm.sup.2 flasks.
190 .mu.l cell suspension was added (at 1.times.10.sup.4) to each
well of columns 2 to 11 of a 96 well plate (Nunc 96F flat bottom TC
plate). 190 .mu.l of media was added to each well of columns 1 and
12. The media was Dulbecco's Modified Eagles' Media (DMEM) (which
included .about.10% Foetal Calf Serum (FCS) and .about.1% 200 mM
L-Glutamine solution).
Plates were incubated overnight at 37.degree. C. before addition of
drug if cells were adherent. 200 .mu.M of the test compound
solutions (in 100% DMSO) were serially diluted across a 96 well
plate. Each resulting point was then further diluted 1/10 into
sterile distilled water (SDW).
To the cell negative blanks and compound negative control wells,
10% DMSO was added at 5% v/v. Assay plates were incubated for the
following durations at 37.degree. C. in 5% CO.sub.2 in a humidified
incubator for 72 hours. Following incubation, MTT solution to a
final concentration of 1.5 .mu.M was added to each well. Plates
were then incubated for a further 4 hours at 37.degree. C. in 5%
CO.sub.2 in a humidified incubator. The media was then removed, and
the dye was solubilised in 200 .mu.l DMSO (99.99%).
Plates were read at 540 nm absorbance using an Envision plate
reader. Data was analysed using Microsoft Excel and GraphPad Prism
and IC.sub.50 values obtained.
Compound II has an IC.sub.50 of 11.7 .mu.M in this assay.
* * * * *
References